Simultaneous red/green dual fluorescence detection on electroblots using BODIPY TR-X succinimidyl ester and ELF 39 phosphate.
AuthorsMartin K, Hart C, Schulenberg B, Jones L, Patton WF
JournalProteomics
PubMed ID11987124
'A two-color fluorescence detection method is described based upon covalently coupling the succinimidyl ester of BODIPY TR-X dye to proteins immobilized on polyvinylidene difluoride membranes, followed by detection of target proteins using the fluorogenic, precipitating substrate ELF 39-phosphate in combination with alkaline phosphatase conjugated reporter molecules. This results in all ... More
Multiphoton confocal microscopy using a femtosecond Cr:forsterite laser.
AuthorsLiu TM, Chu SW, Sun CK, Lin BL, Cheng PC, Johnson I
JournalScanning
PubMed ID11534811
'With its output wavelength covering the infrared penetrating window of most biological tissues at 1,200-1,250 nm, the femtosecond Cr:forsterite laser shows high potential to serve as an excellent excitation source for the multiphoton fluorescence microscope. Its high output power, short optical pulse width, high stability, and low dispersion in fibers ... More
Single-molecule detection of specific nucleic acid sequences in unamplified genomic DNA.
AuthorsCastro A, Williams JG
JournalAnal Chem
PubMed ID9322430
'A new technique is described for the rapid detection of specific nucleic acid sequences in unamplified DNA samples. The method consists of using two nucleic acid probes complementary to different sites on a target DNA sequence. The two probes are each labeled with different fluorescent dyes. When mixed with a ... More
Protein-free parallel triple-stranded DNA complex formation.
'A 14 nt DNA sequence 5''-AGAATGTGGCAAAG-3'' from the zinc finger repeat of the human KRAB zinc finger protein gene ZNF91 bearing the intercalator 2-methoxy,6-chloro,9-amino acridine (Acr) attached to the sugar-phosphate backbone in various positions has been shown to form a specific triple helix (triplex) with a 16 bp hairpin (intramolecular) ... More
Rapid determination of single base mismatch mutations in DNA hybrids by direct electric field control.
AuthorsSosnowski RG, Tu E, Butler WF, O'Connell JP, Heller MJ
JournalProc Natl Acad Sci U S A
PubMed ID9037016
'We have demonstrated that controlled electric fields can be used to regulate transport, concentration, hybridization, and denaturation of single- and double-stranded oligonucleotides. Discrimination among oligonucleotide hybrids with widely varying binding strengths may be attained by simple adjustment of the electric field strength. When this approach is used, electric field denaturation ... More
Simultaneous assay of Src SH3 and SH2 domain binding using different wavelength fluorescence polarization probes.
AuthorsLynch BA, Minor C, Loiacono KA, van Schravendijk MR, Ram MK, Sundaramoorthi R, Adams SE, Phillips T, Holt D, Rickles RJ, MacNeil IA
JournalAnal Biochem
PubMed ID10542110
'pp60(c-src) is a prototypical nonreceptor tyrosine kinase and may play a role in diseases as diverse as cancer and osteoporosis. In Src, the SH3 domain (Src homology 3) binds proteins at specific, proline-rich sequences, while the SH2 domain (Src homology 2) binds phosphotyrosine-containing sequences. Inhibition of Src SH3 and SH2 ... More
Multicolor in vitro translation.
AuthorsTraverso G, Diehl F, Hurst R, Shuber A, Whitney D, Johnson C, Levin B, Kinzler KW, Vogelstein B
JournalNat Biotechnol
PubMed ID12910244
'In vitro translation is a widely used tool for both analytical and preparative purposes. For analytical purposes, small amounts of proteins are synthesized and visualized by detection of labeled amino acids incorporated during translation. The original strategy of incorporating radioactively labeled amino acids, such as [35S]methionine or [14C]leucine, has been ... More
Fraying and electron autodetachment dynamics of trapped gas phase oligonucleotides.
AuthorsDanell AS, Parks JH
JournalJ Am Soc Mass Spectrom
PubMed ID14652182
Sensitive methods recently developed to measure laser-induced fluorescence from trapped ions have been applied to study the dynamics of double- and single-stranded oligonucleotides. In this paper, the fraying of duplex terminal base pairs has been identified by measuring the donor fluorescence as a function of temperature from an oligonucleotide duplex ... More
Coupling immunoassays with chromatographic separation techniques.
AuthorsTang Z, Karnes HT
JournalBiomed Chromatogr
PubMed ID11002281
Coupling immunoassays with HPLC separation techniques is becoming increasingly useful in the analysis of biological and nonbiological samples of both large and small molecules. This is because it provides both sensitivity and selectivity for molecular analysis at relatively low cost, low maintenance and with excellent potential for automation. This paper ... More
Direct, ultrasensitive, and selective optical detection of protein toxins using multivalent interactions.
AuthorsSong X, Swanson BI
JournalAnal Chem
PubMed ID10366891
Three highly sensitive, selective, and reagent-free optical signal transduction methods for detection of polyvalent proteins have been developed by directly coupling distance-dependent fluorescence self-quenching and/or resonant-energy transfer to the protein-receptor binding events. The ganglioside GM1, as the recognition unit for cholera toxin (CT), was covalently labeled with fluorophores and then ... More
Effect of hydrophobic surfactant protein SP-C on binary phospholipid monolayers. Molecular machinery at the air/water interface.
AuthorsKrüger P, Baatz JE, Dluhy RA, Lösche M
JournalBiophys Chem
PubMed ID12408937
Fluorescent and modified dark-field microscopies were used to investigate the phase behavior of physiologically relevant lipid/protein monomolecular films containing surfactant protein C(SP-C). Synthetic human SP-C(1-34) was labeled at its N-terminus using the fluorescent probe 6-(((4(4,4-difluoro-5-(2-thienyl)-4-bora-3a,4a-diaza-s-indacene-3-yl)phenoxy)acetyl)amino)hexanoic acid (BODIPY/TR-X). Using dual fluorescent labeling (lipid and protein) in the monolayers, we have correlated ... More