LYVE-1, a new homologue of the CD44 glycoprotein, is a lymph-specific receptor for hyaluronan.
AuthorsBanerji S, Ni J, Wang SX, Clasper S, Su J, Tammi R, Jones M, Jackson DG
JournalJ Cell Biol
PubMed ID10037799
'The extracellular matrix glycosaminoglycan hyaluronan (HA) is an abundant component of skin and mesenchymal tissues where it facilitates cell migration during wound healing, inflammation, and embryonic morphogenesis. Both during normal tissue homeostasis and particularly after tissue injury, HA is mobilized from these sites through lymphatic vessels to the lymph nodes ... More
Binding of the isolectin I-B4 from Griffonia simplicifolia to the general visceral afferents in the vagus nerve: a light- and electron-microscope study in the rat.
AuthorsLi H, Nomura S, Mizuno N
JournalNeurosci Lett
PubMed ID9121722
'Prominent binding to the isolectin I-B4 from Griffonia simplicifolia (I-B4) was observed not only in the terminal area of general somatic afferents (lamina II of the medullary and the spinal dorsal horns), but also in the terminal area of the general visceral afferents (nucleus of the solitary tract, NST). The ... More
Mps1 is a kinetochore-associated kinase essential for the vertebrate mitotic checkpoint.
AuthorsAbrieu A, Magnaghi-Jaulin L, Kahana JA, Peter M, Castro A, Vigneron S, Lorca T, Cleveland DW, Labbé JC
JournalCell
PubMed ID11461704
'The mitotic checkpoint acts to inhibit entry into anaphase until all chromosomes have successfully attached to spindle microtubules. Unattached kinetochores are believed to release an activated form of Mad2 that inhibits APC/C-dependent ubiquitination and subsequent proteolysis of components needed for anaphase onset. Using Xenopus egg extracts, a vertebrate homolog of ... More
Hu protein as an early marker of neuronal phenotypic differentiation by subependymal zone cells of the adult songbird forebrain.
AuthorsBarami K, Iversen K, Furneaux H, Goldman SA
JournalJ Neurobiol
PubMed ID8586967
'The avian forebrain exhibits neurogenesis in adulthood, with neuronal production from ependymal/subependymal zone (SZ) precursor cells. To follow the commitment of newborn cells to neuronal lineage, we used their expression of the Hu family of neuronal RNA-binding proteins to identify them before their migration from the SZ. Adult canaries were ... More
EphB/syndecan-2 signaling in dendritic spine morphogenesis.
'We previously reported that the cell surface proteoglycan syndecan-2 can induce dendritic spine formation in hippocampal neurons. We demonstrate here that the EphB2 receptor tyrosine kinase phosphorylates syndecan-2 and that this phosphorylation event is crucial for syndecan-2 clustering and spine formation. Syndecan-2 is tyrosine phosphorylated and forms a complex with ... More
High-resolution mapping of human chromosome 11 by in situ hybridization with cosmid clones.
'Cosmid clones containing human DNA inserts have been mapped on chromosome 11 by fluorescence in situ hybridization under conditions that suppress signal from repetitive DNA sequences. Thirteen known genes, one chromosome 11-specific DNA repeat, and 36 random clones were analyzed. High-resolution mapping was facilitated by using digital imaging microscopy and ... More
Fluorescence in situ hybridization (FISH): DNA probe production and hybridization criteria.
AuthorsNath J, Johnson KL
JournalBiotech Histochem
PubMed ID9554580
'We describe methods for the production of fluorescence in situ hybridization (FISH) probes and the utilization of these probes for the detection of complementary DNA sequences with accuracy and sensitivity for application in both basic research and clinical diagnosis. Due to the frequent use of FISH in many laboratories, it ... More
Flow cytometric quantification of human chromosome specific repetitive DNA sequences by single and bicolor fluorescent in situ hybridization to lymphocyte interphase nuclei.
'Fluorescent in situ hybridization allows for rapid and precise detection of specific nucleic acid sequences in interphase and metaphase cells. We applied fluorescent in situ hybridization to human lymphocyte interphase nuclei in suspension to determine differences in amounts of chromosome specific target sequences amongst individuals by dual beam flow cytometry. ... More
Use of methacrylate De-embedding protocols for in situ hybridization on semithin plastic sections with multiple detection strategies.
AuthorsWarren KC, Coyne KJ, Waite JH, Cary SC
JournalJ Histochem Cytochem
PubMed ID9446821
'New plastic resins are gradually replacing traditional paraffin-embedding in situ hybridization (ISH) strategies. One unique resin that has not been fully investigated or exploited with respect to light microscopic ISH is a methacrylate mixture. The advantage of this resin is its ability to be removed from tissues postsectioning, dramatically increasing ... More
Further characterisation of the in situ terminal deoxynucleotidyl transferase (TdT) assay for the flow cytometric analysis of apoptosis in drug resistant and drug sensitive leukaemic cells.
AuthorsChapman RS, Chresta CM, Herberg AA, Beere HM, Heer S, Whetton AD, Hickman JA, Dive C
JournalCytometry
PubMed ID7587710
'Apoptosis, originally defined by specific morphological changes, is characterised biochemically by non-random cleavage of DNA. Depending on cell type, this DNA cleavage proceeds from 300 and 50kbp fragments prior to, concomitantly with, or in the absence of 180bp integer fragmentation. Incorporation into fragmented DNA of biotin-labelled nucleotides by terminal deoxynucleotidyl ... More
Modulation of TGF-beta type 1 receptor: flow cytometric detection with biotinylated TGF-beta.
AuthorsNewman W, Beall LD, Bertolini DR, Cone JL
JournalJ Cell Physiol
PubMed ID2550480
'Transforming growth factor beta type 1 (TGF-beta 1) was reacted with NHS-biotin to yield a derivative of TGF-beta 1 which was biotinylated on lysine residues. The biotinylated form of TGF-beta 1 was separated from the unreacted material by reverse phase chromatography. In three separate bioassays, the derivatized peptide was as ... More
Slow and fast myosin heavy chain content defines three types of myotubes in early muscle cell cultures.
AuthorsMiller JB, Crow MT, Stockdale FE
JournalJ Cell Biol
PubMed ID3902852
'We prepared monoclonal antibodies specific for fast or slow classes of myosin heavy chain isoforms in the chicken and used them to probe myosin expression in cultures of myotubes derived from embryonic chicken myoblasts. Myosin heavy chain expression was assayed by gel electrophoresis and immunoblotting of extracted myosin and by ... More
Biotinylated granulocyte/macrophage colony-stimulating factor analogues: effect of linkage chemistry on activity and binding.
'Biotinylated granulocyte/macrophage colony-stimulating factor (GM-CSF) analogues with different linkage chemistries and levels of conjugated biotin were synthesized by reacting recombinant human GM-CSF with sulfosuccinimidyl 6-biotinamidohexanoate or biotin hydrazide/1-[3-(dimethylamino)-propyl]-3-ethylcarbodiimide. These chemically reactive forms of biotin produced derivatives biotinylated at amine or carboxyl groups, respectively. Amine-derivatized analogues of 1.2 and 3.8 mol ... More
Regulation of the Fas death pathway by FLICE-inhibitory protein in primary human B cells.
AuthorsHennino A, Berard M, Casamayor-Pallejà M, Krammer PH, Defrance T
JournalJ Immunol
PubMed ID10975811
'The Fas/Fas ligand (L) system plays an important role in the maintenance of peripheral B cell tolerance and the prevention of misguided T cell help. CD40-derived signals are required to induce Fas expression on virgin B cells and to promote their susceptibility to Fas-mediated apoptosis. In the current study, we ... More
Selective malformation of the splenic white pulp border in L1-deficient mice.
AuthorsWang SL, Kutsche M, DiSciullo G, Schachner M, Bogen SA
JournalJ Immunol
PubMed ID10946272
'Lymphocytes enter the splenic white pulp by crossing the poorly characterized boundary of the marginal sinus. In this study, we describe the importance of L1, an adhesion molecule of the Ig superfamily, for marginal sinus integrity. We find that germline insertional mutation of L1 is associated with a selective malformation ... More
Fas/Apo-1 (CD95) receptor lacking the intracytoplasmic signaling domain protects tumor cells from Fas-mediated apoptosis.
AuthorsCascino I, Papoff G, De Maria R, Testi R, Ruberti G
JournalJ Immunol
PubMed ID8598453
'FAS/Apo-1 (CD95) is an apoptosis-signaling cell surface receptor belonging to the TNF receptor family. Tumor cells resistant to Fas-mediated apoptosis have been described, but to date, the mechanisms responsible for this resistance are not well understood. We found that a series of apoptosis-resistant clones from human HUT78 lymphoma cells express ... More
Alteration of intracellular calcium flux and impairment of nuclear factor-AT translocation in T cells during acute Toxoplasma gondii infection in mice.
AuthorsHaque S, Dumon H, Haque A, Kasper LH
JournalJ Immunol
PubMed ID9862712
'Down-regulation of host immune response to Toxoplasma gondii is associated with the expression of specific cytokines, in particular IL-10, and the induction of CD4+ T cell anergy. In the present study we report that the expression of both CD4 and CD2 antigen is down-regulated during the acute phase of infection. ... More
Comparative genomic hybridization for molecular cytogenetic analysis of solid tumors.
AuthorsKallioniemi A, Kallioniemi OP, Sudar D, Rutovitz D, Gray JW, Waldman F, Pinkel D
JournalScience
PubMed ID1359641
'Comparative genomic hybridization produces a map of DNA sequence copy number as a function of chromosomal location throughout the entire genome. Differentially labeled test DNA and normal reference DNA are hybridized simultaneously to normal chromosome spreads. The hybridization is detected with two different fluorochromes. Regions of gain or loss of ... More
Localization of unmyelinated axons in rat skin and mucocutaneous tissue utilizing the isolectin GS-I-B4.
AuthorsPetruska JC, Streit WJ, Johnson RD
JournalSomatosens Mot Res
PubMed ID9241725
'The alpha-D-galactose specific isolectin I-B4 from Griffonia simplicifolia (GS-I-B4) labels CNS microglia and certain peripheral neurons, including a subpopulation of small dark, type B dorsal root ganglion cells, some post-ganglionic sympathetic axons, and nearly all peripheral gustatory axons. The innervation patterns of GS-I-B4 reactive sensory ganglion cells are unknown for ... More
Erratic deposition of agrin during the formation of Xenopus neuromuscular junctions in culture.
AuthorsAnderson MJ, Shi ZQ, Grawel R, Zackson SL
JournalDev Biol
PubMed ID7601300
'In order to disclose the mechanism that regulate synapse development we compared the distributions of agrin, acetylcholine receptors (AChR) and a basal lamina heparan sulfate proteoglycan (HSPG) in sections and cultures prepared from Xenopus laevis and Ambystoma mexicanum embryos. While agrin, AChR and HSPG may accumulate almost synchronously at synapses ... More
A versatile means of intracellular labeling: injection of biocytin and its detection with avidin conjugates.
AuthorsHorikawa K, Armstrong WE
JournalJ Neurosci Methods
PubMed ID3146670
'Biocytin is a biotin-lysine complex of low molecular weight containing about 65% biotin, which retains a high affinity for avidin. Since the latter molecule has been conjugated to several histochemical markers, the use of biocytin as an intracellular marker was investigated. Electrodes were filled with a solution of 4-6% biocytin ... More
Anterograde tracing of retinal axons in the avian embryo with low molecular weight derivatives of biotin.
AuthorsHalfter W
JournalDev Biol
PubMed ID2433173
'Several reactive biotin esters were injected into the eyes of chick and quail embryos at various stages of development. Four of the biotin esters reacted with molecules of the eye tissue and were detected with light and electron microscopy in fluorescein isothiocyanate and peroxidase-avidin incubated sections and whole mounts. Intra ... More
Transglutaminase type 1 and its cross-linking activity are concentrated at adherens junctions in simple epithelial cells.
AuthorsHiiragi T, Sasaki H, Nagafuchi A, Sabe H, Shen SC, Matsuki M, Yamanishi K, Tsukita S
JournalJ Biol Chem
PubMed ID10567386
'Transglutaminase type 1 was identified as a tyrosine-phosphorylated protein from the isolated junctional fraction of the mouse liver. This enzyme was reported to be involved in the covalent cross-linking of proteins in keratinocytes, but its expression and activity in other cell types have not been examined. Northern blotting revealed that ... More
Surface membrane biotinylation efficiently mediates the endocytosis of avidin bioconjugates into nucleated cells.
AuthorsWojda U, Goldsmith P, Miller JL
JournalBioconjug Chem
PubMed ID10563774
'Here we demonstrate that biotin covalently attached to cell surface obligates existing receptors to endocytose avidin bioconjugates into nucleated cells. Incubation of fluorescein-labeled avidin with biotinylated cell lines resulted in uniform and rapid surface attachment and endocytosis compared with no detectable association of the avidin-conjugated dye with unbiotinylated cells. Uptake ... More
Protein cargo delivery properties of cell-penetrating peptides. A comparative study.
AuthorsSäälik P, Elmquist A, Hansen M, Padari K, Saar K, Viht K, Langel U, Pooga M
JournalBioconjug Chem
PubMed ID15546190
'Application of cell-penetrating peptides for delivering various hydrophilic macromolecules with biological function into cells has gained much attention in recent years. We compared the protein transduction efficiency of four cell-penetrating peptides: penetratin, Tat peptide, transportan, and pVEC and studied the effects of various medium parameters on the uptake. Depletion of ... More
Conjugated avidin binds to mast cell granules.
AuthorsTharp MD, Seelig LL, Tigelaar RE, Bergstresser PR
JournalJ Histochem Cytochem
PubMed ID2578142
'The glycoprotein, avidin, conjugated either to the enzyme horseradish peroxidase, or to the fluorochrome dyes, fluorescein or rhodamine, identifies the granules of mast cells in both tissues and cell suspensions. In the absence of prior fixation, mast cells were not identified with conjugated avidin; however, granules released from these cells ... More
Cytofluorometric study of lectin binding to the keratinocytes of epidermolysis bullosa simplex.
AuthorsHachisuka H, Nomura H, Sakamoto F, Sasai Y
JournalActa Histochem
PubMed ID2127148
'Epidermolysis bullosa (EB) simplex is a congenital disease that has blister formation following minor mechanical trauma to the skin. The least amount of information concerning the pathogenesis is known in this disease. One possibility is that there are structural abnormalities in keratinocytes. In the present study, we report the binding ... More
Order and genomic distances among members of the carcinoembryonic antigen (CEA) gene family determined by fluorescence in situ hybridization.
AuthorsBrandriff BF, Gordon LA, Tynan KT, Olsen AS, Mohrenweiser HW, Fertitta A, Carrano AV, Trask BJ
JournalGenomics
PubMed ID1572650
'Fluorescence in situ hybridization was used to establish the order of, and to estimate genomic distances among, members of the carcinoembryonic antigen (CEA) and pregnancy-specific glycoprotein (PSG) subgroups on chromosome 19. Fluorescence in situ hybridization to metaphase chromosomes localized the PSG subgroup telomeric to the CEA subgroup. Cosmid clones containing ... More
Identification and stimulation by serotonin of intrinsic sensory neurons of the submucosal plexus of the guinea pig gut: activity-induced expression of Fos immunoreactivity.
AuthorsKirchgessner AL, Tamir H, Gershon MD
JournalJ Neurosci
PubMed ID1729436
'The bowel is the only organ of the body in which neural reflexes can be elicited in the absence of input from the brain or spinal cord. This activity is mediated by the enteric nervous system (ENS), which contains primary afferent neurons. Experiments were carried out to locate the primary ... More
Tyramide signal amplification (TSA)-FISH applied to mapping PCR-labeled probes less than 1 kb in size.
AuthorsSchriml LM, Padilla-Nash HM, Coleman A, Moen P, Nash WG, Menninger J, Jones G, Ried T, Dean M
JournalBiotechniques
PubMed ID10489619
'Tyramide signal amplification (TSA)-FISH was used to map one mouse and two human DNA probes of less than 1 kb in size. The two human probes were 319 and 608 bp, and the mouse probe was 855 bp. Probes, made from PCR products, were labeled by incorporating biotin-11-dUTP (human) and ... More
Simultaneous localization of cosmids and chromosome R-banding by fluorescence microscopy: application to regional mapping of human chromosome 11.
AuthorsCherif D, Julier C, Delattre O, Derré J, Lathrop GM, Berger R
JournalProc Natl Acad Sci U S A
PubMed ID2395867
'A technique for nonradioactive in situ hybridization on human metaphase chromosomes has been developed to localize human cosmid clones. The simple procedure using two fluorescent dyes (fluorescein and propidium iodide) allows the simultaneous identification of chromosomal R-bands and hybridization signal in a single screening of the slides. This technique has ... More
Regulation of connexin degradation as a mechanism to increase gap junction assembly and function.
AuthorsMusil LS, Le AC, VanSlyke JK, Roberts LM
JournalJ Biol Chem
PubMed ID10940315
'Connexins, the integral membrane protein constituents of gap junctions, are degraded at a rate (t(12) = 1.5-5 h) much faster than most other cell surface proteins. Although the turnover of connexins has been shown to be sensitive to inhibitors of either the lysosome or of the proteasome, how connexins are ... More
Two- and three-dimensional distributions of opioid receptors on NG108-15 cells visualized with the aid of fluorescence confocal microscopy and anti-idiotypic antibodies.
AuthorsOrnatowska M, Glasel JA
JournalJ Chem Neuroanat
PubMed ID1316761
'Polyclonal anti-idiotypic antibodies previously shown to be specific for mu- and delta-opioid receptor subclasses on rat brain membrane preparations using radioreceptor assays have been labeled with fluorescent conjugates of avidin. The resulting complexes have been used to study the distribution and properties of opioid receptors on living and fixed NG108-15 ... More
Metabolic selection of glycosylation defects in human cells.
AuthorsYarema KJ, Goon S, Bertozzi CR
JournalNat Biotechnol
PubMed ID11385460
'Changes in glycosylation are often associated with disease progression, but the genetic and metabolic basis of these events is rarely understood in detail at a molecular level. We describe a metabolism-based approach to the selection of mutants in glycoconjugate biosynthesis that provides insight into regulatory mechanisms for oligosaccharide expression and ... More
Use of the avidin-biotin complex for the localization of actin and myosin with fluorescence microscopy.
AuthorsHeggeness MH, Ash JF
JournalJ Cell Biol
PubMed ID326797
'A new indirect method for fluorescence localization of proteins making use of the avidin-biotin complex is described. We have prepared both a biotin-modified rabbit heavy meromyosin (BHMM) and a biotin-modified antibody to a smooth muscle myosin. After fixation, cells can be treated with either BHMM, which binds to actin, or ... More
GlcNAc 2-epimerase can serve a catabolic role in sialic acid metabolism.
'Sialic acid is a major determinant of carbohydrate-receptor interactions in many systems pertinent to human health and disease. N-Acetylmannosamine (ManNAc) is the first committed intermediate in the sialic acid biosynthetic pathway; thus, the mechanisms that control intracellular ManNAc levels are important regulators of sialic acid production. UDP-GlcNAc 2-epimerase and GlcNAc ... More
Clinical applications of flow karyotyping in myelocytic leukemia by stimulation of different subpopulations of cells in blood or bone marrow samples.
AuthorsArkesteijn GJ, van Dekken H, Martens AC, Hagenbeek A
JournalCytometry
PubMed ID2307058
'Examples are presented in which normal as well as abnormal chromosome distributions could be obtained from the same individual by means of bivariate flow karyotyping. Selective stimulation of T-lymphocytes obtained by E-rosetting from the blood of a patient with acute myelocytic leukemia resulted in a normal flow karyogram. The specific ... More
Human lymphocytes making rheumatoid factor and antibody to ssDNA belong to Leu-1+ B-cell subset.
AuthorsCasali P, Burastero SE, Nakamura M, Inghirami G, Notkins AL
JournalScience
PubMed ID3105056
'B lymphocytes bearing the Leu-1 cell-surface antigen (Leu-1+), the human equivalent of mouse Ly-1+ B lymphocytes, have been detected in human peripheral blood, but there is little information on their frequency and properties. Analysis by fluorescence-activated cell sorter and double immunofluorescence showed that Leu-1+ B cells are consistently present in ... More
Fluorescence studies on the mechanism of liposome-cell interactions in vitro.
AuthorsSzoka F, Jacobson K, Derzko Z, Papahadjopoulos D
JournalBiochim Biophys Acta
PubMed ID7397162
'Sonicated unilamellar liposomes containing fluorescent lipid analogs or biotinyl phosphatidylethanolamine as a ligand for fluorescein avidin have been used to study the mechanism of interaction of phospholipid vesicles with eucaryotic cells. Microscopy revealed that after short incubations the fluorescence was associated with the cell surface in a punctate as opposed ... More
Ultra-sensitive FISH is a useful tool for studying chronic HIV-1 infection.
AuthorsDeichmann M, Bentz M, Haas R
JournalJ Virol Methods
PubMed ID9128858
'Using an ultra-sensitive fluorescence in situ hybridization (FISH) protocol, integration sites of the human immunodeficiency virus-1 (HIV-1) were mapped in two model cell lines for chronic HIV infection. In the promonocytic cell line U1/HIV-1, the two integrated copies of proviral HIV-DNA were located on the short arm of one copy ... More
Human fur gene encodes a yeast KEX2-like endoprotease that cleaves pro-beta-NGF in vivo.
AuthorsBresnahan PA, Leduc R, Thomas L, Thorner J, Gibson HL, Brake AJ, Barr PJ, Thomas G
JournalJ Cell Biol
PubMed ID2269657
'Extracts from BSC-40 cells infected with vaccinia recombinants expressing either the yeast KEX2 prohormone endoprotease or a human structural homologue (fur gene product) contained an elevated level of a membrane-associated endoproteolytic activity that could cleave at pairs of basic amino acids (-LysArg- and -ArgArg-). The fur-directed activity (furin) shared many ... More
Specific lysis of human tumor cells by T cells coated with anti-T3 cross-linked to anti-tumor antibody.
AuthorsPerez P, Titus JA, Lotze MT, Cuttitta F, Longo DL, Groves ES, Rabin H, Durda PJ, Segal DM
JournalJ Immunol
PubMed ID2944946
'Heteroaggregates containing anti-T3 cross-linked to anti-target cell antibodies have been shown to cause human T cells to lyse target cells that express antigens recognized by the anti-target cell antibody. In this study, we test targeted human T cells for the ability to lyse human tumor cells as a first step ... More
Spatial topography of a pericentromeric region (1q12) in hemopoietic cells studied by in situ hybridization and confocal microscopy.
Authorsvan Dekken H, van Rotterdam A, Jonker RR, van der Voort HT, Brakenhoff GJ, Bauman JG
JournalCytometry
PubMed ID2379447
'A fluorescent in situ hybridization procedure with a chromosome 1-specific (1q12) repetitive satellite DNA probe was used to label the 1q12 regions of the chromosomes 1 in spherical and polymorphic hemopoietic cell nuclei. The entire procedure was performed in suspension to preserve nuclear morphology. The result was studied by three-dimensional ... More
Fluorimetric assays for avidin and biotin based on biotin-induced fluorescence enhancement of fluorescein-labeled avidin.
AuthorsAl-Hakiem MH, Landon J, Smith DS, Nargessi RD
JournalAnal Biochem
PubMed ID6797319
BAC mapping using fluorescence in situ hybridization.
AuthorsChen XN, Korenberg JR
JournalMethods Mol Biol
PubMed ID15020820
Cytogenetic analysis by in situ hybridization with fluorescently labeled nucleic acid probes.
AuthorsPinkel D, Gray JW, Trask B, van den Engh G, Fuscoe J, van Dekken H
JournalCold Spring Harb Symp Quant Biol
PubMed ID3472711
Approaches to multicolor immunofluorescence measurements.
AuthorsHoffman RA
JournalCytometry Suppl
PubMed ID3076363
Fluorescent localization of contractile proteins in tissue culture cells.
AuthorsWang K, Feramisco JR, Ash JF
JournalMethods Enzymol
PubMed ID6750319
Amplification of the biotin-avidin immunmofluorescence technique.
AuthorsBerman JW, Basch RS
JournalJ Immunol Methods
PubMed ID7000912
An amplification of the immunofluorescence technique which uses biotinylated antibody and fluoresceinated avidin is described. By introducing a sandwich technique using fluorescein-conjugated goat anti-avidin, a 5-fold enhancement of staining over the conventional immunofluorescence method is achieved, and the brightness is more than twice that achieved with the simple biotin-fluoresceinated avidin ... More
Single-molecule fluorescence observed with mercury lamp illumination.
AuthorsUnger M, Kartalov E, Chiu CS, Lester HA, Quake SR
JournalBiotechniques
PubMed ID10572649
We demonstrate that it is possible to observe single fluorescent molecules using a standard fluorescence microscope with mercury lamp excitation and an inexpensive cooled charge-coupled device (CCD) camera. With this equipment, we have been able to observe single molecules of tetramethyl-rhodamine, rhodamine 6G, fluorescein isothiocyanate and green fluorescent protein. Immobilized ... More
A simple method for immunoselective cell separation with the avidin-biotin system.
AuthorsWormmeester J, Stiekema F, De Groot K
JournalJ Immunol Methods
PubMed ID6368688
A new method for immunoselective cell separation is described which uses biotinylated antibodies and avidin-coupled sheep erythrocytes for rosette formation, and a density gradient to separate the rosetting from the non-rosetting cells. The utility of the technique is illustrated by separating subpopulations of rabbit, mouse and rat lymphocytes. The method ... More
"DAKLI": a multipurpose ligand with high affinity and selectivity for dynorphin (kappa opioid) binding sites.
AuthorsGoldstein A, Nestor JJ, Naidu A, Newman SR
JournalProc Natl Acad Sci U S A
PubMed ID2902630
We describe a synthetic ligand, "DAKLI" (Dynorphin A-analogue Kappa LIgand), related to the opioid peptide dynorphin A. A single reactive amino group at the extended carboxyl terminus permits various reporter groups to be attached, such as 125I-labeled Bolton-Hunter reagent, fluorescein isothiocyanate, or biotin. These derivatives have high affinity and selectivity ... More
Direct nonradioactive in situ hybridization of somatic cell hybrid DNA to human lymphocyte chromosomes.
Biotinylated DNA from various human-rodent hybrids was hybridized to human lymphocyte spreads after preannealing of the repeated sequences with sonicated total human DNA. Fluorescent labeling was achieved by successive treatments with fluorescein-labeled avidin and biotinylated antiavidin antibody. The use of labeled total DNA from hybrids with known chromosome composition permits ... More
Direct effects of radiation on the avidin-biotin system. Absence of energy transfer.
AuthorsKempner ES, Miller JH
JournalJ Biol Chem
PubMed ID2203786
Frozen solutions of biotinylated glucose-6-phosphate dehydrogenase and fluorescently tagged avidin were exposed to high energy ionizing radiation. Parallel experiments with peroxidase coupled to streptavidin and with biotinylated phycoerythrin were also performed. The loss of function of each compound was analyzed according to target theory. Target analysis revealed that the radiation-sensitive ... More
Three-dimensional reconstruction of pericentromeric (1q12) DNA and ribosomal RNA sequences in HL60 cells after double-target in situ hybridization and confocal microscopy.
Authorsvan Dekken H, van der Voort HT, Brakenhoff GJ, Bauman JG
JournalCytometry
PubMed ID2379448
A fluorescent in situ hybridization procedure was applied to simultaneously label intranuclear pericentromeric (1q12) sequences of the chromosomes 1 and cytoplasmic ribosomal RNA sequences in whole cells of the promyelocytic HL60 cell line. For this purpose biotinated chromosome 1-specific (1q12) repetitive satellite DNA and 28S ribosomal ssRNA probes were used. ... More
Membrane sialoglycolipids emerging as possible signal transducers for lymphocyte stimulation.
AuthorsSpiegel S, Wilchek M
JournalJ Immunol
PubMed ID6972969
Biotin hydrazide was attached covalently to the aldehyde groups produced by periodate oxidation of bovine brain gangliosides. These modified gangliosides were incorporated into mature rat thymocytes by incubation of the biotinyl gangliosides in the culture medium containing these cells. Avidin, which binds strongly to biotin, agglutinated and stimulated DNA synthesis ... More
A rapid staining procedure for two-color analysis of lymphocyte antigen expression.
AuthorsPechhold K, Kabelitz D
JournalJ Immunol Methods
PubMed ID1371796
Two color immunofluorescence analysis of lymphocyte cell surface antigen expression using an unconjugated plus a biotinylated monoclonal antibody (mAb) requires four incubation steps: (1) unconjugated mAb; (2) fluorochrome-labelled goat anti-mouse Ig; (3) biotinylated mAb; (4) fluorochrome-labelled avidin or streptavidin. We describe a time-saving modification of this procedure which requires only ... More
Detection of cytomegalovirus by DNA-DNA hybridization employing probes labelled with 32-phosphorus or biotin.
Various factors influencing the detection of human cytomegalovirus (HCMV) in infected cells by DNA-DNA hybridization have been investigated. Employing the Hind III O fragment of HCMV AD169 labelled with 32P, we found that detection sensitivity was highly influenced by the method employed for extraction of DNA from infected cells. Excision ... More
Survival and size are differentially regulated by placental and fetal PKBalpha/AKT1 in mice.
AuthorsPlaks V, Berkovitz E, Vandoorne K, Berkutzki T, Damari GM, Haffner R, Dekel N, Hemmings BA, Neeman M, Harmelin A,
JournalBiol Reprod
PubMed ID20980686
The importance of placental circulation is exemplified by the correlation of placental size and blood flow with fetal weight and survival during normal and compromised human pregnancies in such conditions as preeclampsia and intrauterine growth restriction (IUGR). Using noninvasive magnetic resonance imaging, we evaluated the role of PKBalpha/AKT1, a major ... More
Advanced glycation end products induce a prothrombotic phenotype in mice via interaction with platelet CD36.
AuthorsZhu W, Li W, Silverstein RL,
JournalBlood
PubMed ID22431576
Diabetes mellitus has been associated with platelet hyperreactivity, which plays a central role in the hyperglycemia-related prothrombotic phenotype. The mechanisms responsible for this phenomenon are not established. In the present study, we investigated the role of CD36, a class-B scavenger receptor, in this process. Using both in vitro and in ... More
Dystrophin is a component of the subsynaptic membrane.
AuthorsYeadon JE, Lin H, Dyer SM, Burden SJ
JournalJ Cell Biol
PubMed ID1720119
A subsynaptic protein of Mr approximately 300 kD is a major component of Torpedo electric organ postsynaptic membranes and copurifies with the AChR and the 43-kD subsynaptic protein. mAbs against this protein react with neuromuscular synapses in higher vertebrates, but not at synapses in dystrophic muscle. The Torpedo 300-kD protein ... More
Analysis of protein binding to receptor-doped lipid monolayers by Monte Carlo simulation.
AuthorsZhao S, Reichert WM
JournalBiophys J
PubMed ID8161683
This paper presents a Monte Carlo simulation (MCS) method for estimating the parameters that characterize ligand-receptor binding directly from experimentally derived binding isotherms. Binding parameters are estimated by incorporating an MCS algorithm for ligand binding to a two-dimensional receptor array into a nonlinear regression program. The MCS method was tested ... More
Detection of biotinylated proteins in polyacrylamide gels using an avidin-fluorescein conjugate.
AuthorsNakamura M, Tsumoto K, Ishimura K, Kumagai I
JournalAnal Biochem
PubMed ID12009700
Biotinylated proteins are widely used as a molecular tool in biotechnological applications. In this paper, we demonstrated that biotinylated proteins after electrophoresis were detected directly in gels using an avidin-fluorescein conjugate with a fluorescence image analyzer. Upon analysis of the purified and chemically biotinylated protein, the sensitivity of this method ... More
Enzymatic production of single-stranded DNA as a target for fluorescence in situ hybridization.
This study demonstrates that Exonuclease III (Exo III) can be used to produce sufficient single-stranded (ss)DNA in chromosomes and cells to allow in situ hybridization. In this study, all of the probes were modified with biotin and the probe binding was visualized with fluorescein-labeled avidin. Exo III digestion starting at ... More
Interphase and metaphase resolution of different distances within the human dystrophin gene.
AuthorsLawrence JB, Singer RH, McNeil JA
JournalScience
PubMed ID2203143
Fluorescence in situ hybridization makes possible direct visualization of single sequences not only on chromosomes, but within decondensed interphase nuclei, providing a potentially powerful approach for high-resolution (1 Mb and below) gene mapping and the analysis of nuclear organization. Interphase mapping was able to extend the ability to resolve and ... More
Focal sites of DNA repair synthesis in human chromosomes.
Nucleotide excision repair (NER) is the principle pathway by which the human cells eliminate UV-induced lesions from their genomic DNA. The process can be visualized through the labelling of the nucleotides that are incoporated into repair patches, following the excision of the damaged stretch of DNA. In this study we ... More
Cytogenetic analysis using quantitative, high-sensitivity, fluorescence hybridization.
AuthorsPinkel D, Straume T, Gray JW
JournalProc Natl Acad Sci U S A
PubMed ID3458254
This report describes the use of fluorescence in situ hybridization for chromosome classification and detection of chromosome aberrations. Biotin-labeled DNA was hybridized to target chromosomes and subsequently rendered fluorescent by successive treatments with fluorescein-labeled avidin and biotinylated anti-avidin antibody. Human chromosomes in human-hamster hybrid cell lines were intensely and uniformly ... More
Rac1b and reactive oxygen species mediate MMP-3-induced EMT and genomic instability.
AuthorsRadisky DC, Levy DD, Littlepage LE, Liu H, Nelson CM, Fata JE, Leake D, Godden EL, Albertson DG, Nieto MA, Werb Z, Bissell MJ
JournalNature
PubMed ID16001073
The tumour microenvironment can be a potent carcinogen, not only by facilitating cancer progression and activating dormant cancer cells, but also by stimulating tumour formation. We have previously investigated stromelysin-1/matrix metalloproteinase-3 (MMP-3), a stromal enzyme upregulated in many breast tumours, and found that MMP-3 can cause epithelial-mesenchymal transition (EMT) and ... More
Actin-binding specificity of marine macrolide toxins, mycalolide B and kabiramide D.
AuthorsWada S, Matsunaga S, Saito S, Fusetani N, Watabe S
JournalJ Biochem (Tokyo)
PubMed ID9562630
An actin-depolymerizing marine natural product, mycalolide B, and a related compound, kabiramide D, were labeled with biocytin, a biotin derivative, and used to specify target molecules in cultured rat 3Y1 fibroblasts. Mycalolide B exhibited the ability to bind to various intracellular proteins, probably through the Michael addition of a sulfhydryl ... More
Actin motion on microlithographically functionalized myosin surfaces and tracks.
AuthorsNicolau DV, Suzuki H, Mashiko S, Taguchi T, Yoshikawa S
JournalBiophys J
PubMed ID10423457
High-resolution e-beam patterning exposure of the surface of poly[(tert-butyl-methacrylate)-co-(methyl methacrylate)]-a common e-beam and deep-UV resist used in semiconductor microlithography-induced sharp changes in the surface hydrophobicity. These differences in hydrophobicity resulted in the selective attachment of heavy meromyosin to hydrophobic, unexposed surfaces. The movement of the actin filaments on myosin-rich and ... More
Mapping chromosome band 11q23 in human acute leukemia with biotinylated probes: identification of 11q23 translocation breakpoints with a yeast artificial chromosome.
Translocations involving chromosome 11, band q23, are frequent recurring abnormalities in human acute lymphoblastic and acute myeloid leukemia. We used 19 biotin-labeled probes derived from genes and anonymous cosmids for hybridization to metaphase chromosomes from leukemia cells that contained four translocations involving band 11q23: t(4;11)(q21;q23), t(6;11)(q27;q23), t(9;11)(p22;q23), and t(11;19)(q23;p13). The ... More
Visualization of oligonucleotide probes and point mutations in interphase nuclei and DNA fibers using rolling circle DNA amplification.
AuthorsZhong XB, Lizardi PM, Huang XH, Bray-Ward PL, Ward DC
JournalProc Natl Acad Sci U S A
PubMed ID11274414
Rolling circle amplification (RCA) is a surface-anchored DNA replication reaction that can be exploited to visualize single molecular recognition events. Here we report the use of RCA to visualize target DNA sequences as small as 50 nts in peripheral blood lymphocytes or in stretched DNA fibers. Three unique target sequences ... More
Fluorescence in situ hybridization with Alu and L1 polymerase chain reaction probes for rapid characterization of human chromosomes in hybrid cell lines.
AuthorsLichter P, Ledbetter SA, Ledbetter DH, Ward DC
JournalProc Natl Acad Sci U S A
PubMed ID2395866
Human-rodent hybrid cell lines have been analyzed with regard to their human DNA content by using various DNA probe sets, derived from the hybrids, for in situ hybridization to normal human metaphase chromosome spreads. Total genomic hybrid DNA was compared with probe sets of hybrid DNA that were highly enriched ... More
Localization of a mouse centromeric DNA repeat in interphase nuclei.
AuthorsDisteche CM, Adler DA
JournalCytometry
PubMed ID2307052
The position of a mouse DNA repeat located near the centromere of mouse chromosomes X, 11, 13, and 17 was examined in interphase nuclei of bone marrow and fibroblast cells by in situ hybridization of 3H- or biotin-labeled DNA probe 70-38. In most laboratory mouse strains this probe recognizes a ... More
The coordinate organization of vinculin and of actin filaments during the early stages of fibroblast spreading on a substratum.
AuthorsDavid-Pfeuty T
JournalEur J Cell Biol
PubMed ID3922762
Cultured normal fibroblasts adhere to their support essentially through the focal adhesion plaques which are greatly enriched with the 130 000 dalton protein, vinculin, along with the newly described 215 000 dalton protein, talin, and at which actin bundles terminate. In order to explore a role for vinculin in the ... More
Histamine treatment induces rearrangements of orthogonal arrays of particles (OAPs) in human AQP4-expressing gastric cells.
AuthorsCarmosino M, Procino G, Nicchia GP, Mannucci R, Verbavatz JM, Gobin R, Svelto M, Valenti G
JournalJ Cell Biol
PubMed ID11564760
To test the involvement of the water channel aquaporin (AQP)-4 in gastric acid physiology, the human gastric cell line (HGT)-1 was stably transfected with rat AQP4. AQP4 was immunolocalized to the basolateral membrane of transfected HGT-1 cells, like in native parietal cells. Expression of AQP4 in transfected cells increased the ... More
Use of fluorescent sequence-specific polyamides to discriminate human chromosomes by microscopy and flow cytometry.
In this paper, we demonstrate the use of synthetic polyamide probes to fluorescently label heterochromatic regions on human chromosomes for discrimination in cytogenetic preparations and by flow cytometry. Polyamides bind to the minor groove of DNA in a sequence-specific manner. Unlike conventional sequence-specific DNA or RNA probes, polyamides can recognize ... More
Calcium-dependent conformation of a mouse macrophage calcium-type lectin. Carbohydrate binding activity is stabilized by an antibody specific for a calcium-dependent epitope.
AuthorsKimura T, Imai Y, Irimura T
JournalJ Biol Chem
PubMed ID7541793
We established monoclonal antibodies (mAbs) against the mouse macrophage galactose/N-acetylgalactosamine-specific lectin (MMGL) that is a 42-kDa calcium-dependent lectin, using a solid phase carbohydrate binding assay as a novel strategy for screening mAbs. The specificity of six mAbs were investigated by antibody binding to native or recombinant forms (rML) of MMGL, ... More
Coupling of biotin-(poly(ethylene glycol))amine to poly(D,L-lactide-co-glycolide) nanoparticles for versatile surface modification.
AuthorsWeiss B, Schneider M, Muys L, Taetz S, Neumann D, Schaefer UF, Lehr CM
JournalBioconjug Chem
PubMed ID17590034
Generally, polymeric nanoparticles (NP) for drug targeting are designed to entrap the drug moiety in the core and to present the targeting moiety on the surface. However, in most cases, common preparation techniques of polymeric NP need to be specifically arranged for each compound to be entrapped or attached. In ... More
Flow cytometric analysis of recombinant murine GM-CSF (rmuGM-CSF) induced changes in the distribution of specific cell populations in vivo.
AuthorsPennline KJ, Pellerito F, DaFonseca M, Monahan P, Siegel MI, Smith SR
JournalCytometry
PubMed ID2180653
The changes in the distribution of granulocytes, monocytes, and lymphocytes in various tissue compartments following subcutaneous (SC) administration of recombinant murine GM-CSF (rmuGM-CSF) in vivo was determined by flow cytometry in time course studies. Balb/c mice were given single, daily SC injections of 1 or 4 micrograms of rmuGM-CSF for ... More
Discrete nuclear domains of poly(A) RNA and their relationship to the functional organization of the nucleus.
AuthorsCarter KC, Taneja KL, Lawrence JB
JournalJ Cell Biol
PubMed ID1720123
The functional organization of the nucleus was studied using a fluorescence microscopy approach which allowed integration of positional information for RNA, DNA, and proteins. In cells from sea urchin to human, nuclear poly(A) RNA was found concentrated primarily within several discrete "transcript domains" which often surrounded nucleoli. Concentrations of poly(A) ... More
Rapid metaphase and interphase detection of radiation-induced chromosome aberrations in human lymphocytes by chromosomal suppression in situ hybridization.
AuthorsCremer T, Popp S, Emmerich P, Lichter P, Cremer C
JournalCytometry
PubMed ID2307051
Chromosomal in situ suppression (CISS)-hybridization of biotinylated phage DNA-library inserts from sorted human chromosomes was used to decorate chromosomes 1 and 7 specifically from pter to qter and to detect structural aberrations of these chromosomes in irradiated human peripheral lymphocytes. In addition, probe pUC1.77 was used to mark the 1q12 ... More
Antigen-induced changes in B cell subsets in lymph nodes: analysis by dual fluorescence flow cytofluorometry.
AuthorsKraal G, Hardy RR, Gallatin WM, Weissman IL, Butcher EC
JournalEur J Immunol
PubMed ID2873041
Changes in the representation and surface phenotype of defined B cell subsets in murine lymph nodes stimulated with keyhole limpet hemocyanin or sheep red blood cells have been analyzed by two-color immunofluorescence fluorocytometric analysis. Shortly after immunization with either antigen there is a dramatic increase in both the frequency and ... More
Mapping small DNA sequences by fluorescence in situ hybridization directly on banded metaphase chromosomes.
AuthorsFan YS, Davis LM, Shows TB
JournalProc Natl Acad Sci U S A
PubMed ID2201023
A procedure for mapping small DNA probes directly on banded human chromosomes by fluorescence in situ hybridization has been developed. This procedure allows for the simultaneous visualization of banded chromosomes and hybridization signal without overlaying two separate photographic images. This method is simple and rapid, requires only a typical fluorescence ... More
Purification of gonadotropes and intracellular free calcium oscillation. Effects of gonadotropin-releasing hormone and interleukin 6.
AuthorsMasumoto N, Tasaka K, Kasahara K, Miyake A, Tanizawa O
JournalJ Biol Chem
PubMed ID2007597
The intracellular free calcium concentration ([Ca2+]i) in single gonadotropes was measured with a calcium-sensitive fluorescent dye indo-1 or fura-2 and a digital imaging fluorescence microscopic system to determine how interleukin-6 (IL-6) increases release of gonadotropins. IL-6 induced an increase in the basal [Ca2+]i or the amplitude of spontaneous oscillation of ... More
Conjugated avidin identifies cutaneous rodent and human mast cells.
AuthorsBergstresser PR, Tigelaar RE, Tharp MD
JournalJ Invest Dermatol
PubMed ID6470526
Avidin conjugated to the fluorescent dyes rhodamine or fluorescein binds to mast cell granules in rodent and human skin. Sequential staining of tissue mast cells first with conjugated avidin, and then with a metachromatic stain demonstrated that both techniques identify the same mast cell granules. Specificity for mast cells was ... More
B cell activation. VI. Effects of exogenous diglyceride and modulators of phospholipid metabolism suggest a central role for diacylglycerol generation in transmembrane signaling by mIg.
AuthorsCoggeshall KM, Cambier JC
JournalJ Immunol
PubMed ID2981089
Previous evidence indicates that in vitro activators of protein kinase C, such as phorbol myristate acetate (PMA), are able to induce early activation events in murine B cells, including membrane depolarization and increased I-A antigen expression. These same events are induced by specific antigen and anti-receptor antibody. This evidence suggests ... More
Antennapedia and HIV transactivator of transcription (TAT) "protein transduction domains" promote endocytosis of high molecular weight cargo upon binding to cell surface glycosaminoglycans.
AuthorsConsole S, Marty C, García-Echeverría C, Schwendener R, Ballmer-Hofer K
JournalJ Biol Chem
PubMed ID12837762
Protein transduction domains (PTDs) are short basic peptide sequences present in many cellular and viral proteins that mediate translocation across cellular membranes. PTDs have become widely used as tools for the delivery of high Mr polypeptides, polynucleotides, or nanoparticles to cells in culture; and even the transfer of cargo molecules ... More
Analysis and isolation of embryonic mammalian neurons by fluorescence-activated cell sorting.
AuthorsSt John PA, Kell WM, Mazzetta JS, Lange GD, Barker JL
JournalJ Neurosci
PubMed ID2872281
Cells were dissociated from the CNS of the embryonic mouse and rat to produce cell suspensions suitable for analysis and separation on a fluorescence-activated cell sorter (FACS). Cells from the spinal cord of the embryonic mouse were analyzed in the most detail. Cell suspensions generated three major peaks in histograms ... More
Properties and regulation of gap junctional hemichannels in the plasma membranes of cultured cells.
AuthorsLi H, Liu TF, Lazrak A, Peracchia C, Goldberg GS, Lampe PD, Johnson RG
JournalJ Cell Biol
PubMed ID8769424
During the assembly of gap junctions, a hemichannel in the plasma membrane of one cell is thought to align and dock with another in an apposed membrane to form a cell-to-cell channel. We report here on the existence and properties of nonjunctional, plasma membrane connexin43 (Cx43) hemichannels. The opening of ... More
Electric field-induced redistribution and postfield relaxation of low density lipoprotein receptors on cultured human fibroblasts.
AuthorsTank DW, Fredericks WJ, Barak LS, Webb WW
JournalJ Cell Biol
PubMed ID2861206
The lateral mobility of unliganded low density lipoprotein-receptor (LDL-R) on the surface of human fibroblasts has been investigated by studying the generation and relaxation of concentration differences induced by exposure of the cultured cells to steady electric fields. The topographic distribution of receptors was determined by fluorescence microscopy of cells ... More
Phenotype of recovering lymphoid cell populations after marrow transplantation.
AuthorsAult KA, Antin JH, Ginsburg D, Orkin SH, Rappeport JM, Keohan ML, Martin P, Smith BR
JournalJ Exp Med
PubMed ID3159819
Four patients who received bone marrow transplants were studied sequentially during the posttransplant period to define the pattern of recovering lymphoid cell types. Three patients received T cell-depleted, HLA-matched marrow, and one received untreated marrow from an identical twin. Blood lymphoid cells were labeled with 25 different pairs of monoclonal ... More
Tyrosine phosphorylation and acetylcholine receptor cluster formation in cultured Xenopus muscle cells.
AuthorsBaker LP, Peng HB
JournalJ Cell Biol
PubMed ID7678011
Aggregation of the nicotinic acetylcholine receptor (AChR) at sites of nerve-muscle contact is one of the earliest events to occur during the development of the neuromuscular junction. The stimulus presented to the muscle by nerve and the mechanisms underlying postsynaptic differentiation are not known. The purpose of this study was ... More
Characterization of the purified Chlamydomonas minus agglutinin.
AuthorsCollin-Osdoby P, Adair WS
JournalJ Cell Biol
PubMed ID2411736
Chlamydomonas flagellar sexual agglutinins are responsible for the adhesion of opposite mating-type (plus and minus) gametes during the first stages of mating. Purification and partial characterization of the plus agglutinin was previously reported (Adair, W. S., C. J. Hwang, and U. W. Goodenough, 1983, Cell, 33:183-193). Here we characterize the ... More
Probing the ligand-binding domain of the mGluR4 subtype of metabotropic glutamate receptor.
Metabotropic glutamate receptors (mGluRs) are G-protein-coupled glutamate receptors that subserve a number of diverse functions in the central nervous system. The large extracellular amino-terminal domains (ATDs) of mGluRs are homologous to the periplasmic binding proteins in bacteria. In this study, a region in the ATD of the mGluR4 subtype of ... More
Primaquine interferes with membrane recycling from endosomes to the plasma membrane through a direct interaction with endosomes which does not involve neutralisation of endosomal pH nor osmotic swelling of endosomes.
Authorsvan Weert AW, Geuze HJ, Groothuis B, Stoorvogel W
JournalEur J Cell Biol
PubMed ID10928454
The anti-malaria drug primaquine is a weak base which accumulates in endosomes in a protonated form and consequently neutralises the endosomal pH. Bafilomycin A1 prevents endosome acidification by inhibiting the vacuolar proton pump. Although both agents neutralise the endosomal pH, only primaquine has a strong inhibitory effect on recycling of ... More
The use of cationized ferritin to measure cell surface charge of mouse bone marrow cells by flow cytometry.
AuthorsBauman JG, Bouwman E
JournalHistochemistry
PubMed ID3087917
We have prepared fluorescein isothiocyanate (FITC) conjugates of cationised ferritin (CF) and have investigated the usefulness of this CF-FITC to measure the negative cell surface charge of mouse bone marrow cells by flow cytometry. CF-FITC conjugates of low fluorochrome to protein ratios (F/P ratio) gave insufficient fluorescence and/or formed large ... More
Detection of a glycosylation-dependent ligand for the T lymphocyte cell adhesion molecule CD2 using a novel multimeric recombinant CD2-binding assay.
The CD2 molecule plays an important role in T cell adhesion by interacting with the ligands CD58 (LFA-3) and CD59. In order to detect additional ligands for CD2, potentially of low binding affinity, we have prepared a highly fluorescent, multimeric form of rCD2 whose binding to cells can be quantified ... More
Preservation of specific RNA distribution within the chromatin-depleted nuclear substructure demonstrated by in situ hybridization coupled with biochemical fractionation.
AuthorsXing YG, Lawrence JB
JournalJ Cell Biol
PubMed ID1705562
Biochemical fractionation procedures previously shown to remove 95% of cellular protein, DNA, and phospholipid, were combined with fluorescence in situ hybridization to provide a critical evaluation of the retention and spatial preservation of specific primary transcripts within the chromatin-depleted nuclear substructure, operationally defined as the nuclear "matrix." This unique approach ... More