Monoclonal antibodies to NTF2 inhibit nuclear protein import by preventing nuclear translocation of the GTPase Ran.
AuthorsSteggerda SM, Black BE, Paschal BM
JournalMol Biol Cell
PubMed ID10679025
'Nuclear transport factor 2 (NTF2) is a soluble transport protein originally identified by its ability to stimulate nuclear localization signal (NLS)-dependent protein import in digitonin-permeabilized cells. NTF2 has been shown to bind nuclear pore complex proteins and the GDP form of Ran in vitro. Recently, it has been reported that ... More
Development of homogeneous binding assays based on fluorescence resonance energy transfer between quantum dots and Alexa Fluor fluorophores.
AuthorsNikiforov TT, Beechem JM
JournalAnal Biochem
PubMed ID16860286
'We studied the fluorescence resonance energy transfer (FRET) between quantum dots emitting at 565, 605, and 655 nm as energy donors and Alexa Fluor fluorophores with absorbance maxima at 594, 633, 647, and 680 nm as energy acceptors. As a first step, we prepared covalent conjugates between all three types ... More
Preparation and properties of anti-biotin antibodies.
AuthorsKohen F, Bagci H, Barnard G, Bayer EA, Gayer B, Schindler DG, Ainbinder E, Wilchek M
JournalMethods Enzymol
PubMed ID9211297
Monoclonal antibody to biotin.
AuthorsDakshinamurti K, Rector ES
JournalMethods Enzymol
PubMed ID2388565
Monoclonal anti-biotin antibodies simulate avidin in the recognition of biotin.
AuthorsBagçi H, Kohen F, Kusçuoglu U, Bayer EA, Wilchek M
JournalFEBS Lett
PubMed ID8482366
The sequence of the VH gene of a monoclonal anti-biotin antibody was determined. Biotin-binding motifs, similar to those in avidin and streptavidin, were identified in complementary determining regions 2 and 3, suggesting that natural selection of functional motifs may occur in unrelated protein types. ... More
Anti-biotin antibodies offer superior organelle-specific labeling of mitochondria over avidin or streptavidin.
AuthorsHollinshead M, Sanderson J, Vaux DJ
JournalJ Histochem Cytochem
PubMed ID9267466
The mitochondrial matrix contains endogenously biotinylated proteins. These proteins can cause unexpected background signal when biotin-avidin- or biotin-streptavidin-based detection systems are used in immunocytochemistry. Here we show that this reactivity can be deliberately exploited, using a simple anti-biotin reagent, to obtain strong and highly specific labeling of mitochondria by both ... More
Surface plasmon resonance (SPR) as a tool for antibody conjugate analysis.
AuthorsAdamczyk M, Mattingly PG, Shreder K, Yu Z
JournalBioconjug Chem
PubMed ID10563772
Surface plasmon resonance (SPR) analysis was used to assess the immunoreactivity of anti-biotin (4) and anti-fluorescein (5) monoclonal antibody after conjugation with the N-hydroxysuccinimide ester of acridinium-9-carboxamide 1. Only minor changes in the apparent equilibrium dissociation constants of the antibody conjugates for their ligands resulted from the conjugation process. However, ... More
Metabolic biotinylation of cell surface receptors for in vivo imaging.
AuthorsTannous BA, Grimm J, Perry KF, Chen JW, Weissleder R, Breakefield XO
JournalNat Methods
PubMed ID16628210
We have developed a versatile, potent technique for imaging cells in culture and in vivo by expressing a metabolically biotinylated cell-surface receptor and visualizing it with labeled streptavidin moieties. The recombinant reporter protein, which incorporates a biotin acceptor peptide (BAP) between an N-terminal signal sequence and a transmembrane domain, (BAP-TM) ... More
Detection of Cryptosporidium parvum using oligonucleotide-tagged liposomes in a competitive assay format.
AuthorsEsch MB, Baeumner AJ, Durst RA
JournalAnal Chem
PubMed ID11467568
To meet the technical challenge of accurately and rapidly detecting Cryptosporidium parvum oocysts in environmental water, the authors developed a single-use visual-strip assay. The first step in the overall assay procedure involves extracting C. parvum's mRNA coding for heat-shock protein hsp70, followed by amplification using nucleic acid sequence-based amplification (NASBA) ... More
Localization and recycling of gp27 (hp24gamma3): complex formation with other p24 family members.
AuthorsFüllekrug J, Suganuma T, Tang BL, Hong W, Storrie B, Nilsson T
JournalMol Biol Cell
PubMed ID10359607
We report here the characterization of gp27 (hp24gamma3), a glycoprotein of the p24 family of small and abundant transmembrane proteins of the secretory pathway. Immunoelectron and confocal scanning microscopy show that at steady state, gp27 localizes to the cis side of the Golgi apparatus. In addition, some gp27 was detected ... More