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Dot-blot assay for heparin-binding proteins

Journal Article · · Anal. Biochem.; (United States)
; ; ;

A method for the detection and quantitation of picomole amounts of heparin-binding proteins is described. Proteins are first spotted on nitrocellulose and then incubated with /sup 125/I-heparin. Binding of heparin to the proteins is detected by radioautography and quantitated by scanning densitometry; proteins are quantitated by densitometric analysis of the amido black stained nitrocellulose. Heparin-binding was time-dependent and sensitive to the presence of metal ions, urea, and detergents (anionic, nonionic, and zwitterionic). The divalent cations Ca/sup 2 +/ and Mg/sup 2 +/ and the zwitterionic detergent 3-((3-cholamidopropyl)dimethylammonio)-1-propanesulfonate increased heparin binding whereas NaCl, urea, sodium dodecylsulfate, and La3+ decreased binding. This assay is applicable to the identification and characterization of a variety of heparin-binding proteins.

Research Organization:
Merrell Dow Research Institute, Cincinnati, OH
OSTI ID:
7190062
Journal Information:
Anal. Biochem.; (United States), Journal Name: Anal. Biochem.; (United States) Vol. 2; ISSN ANBCA
Country of Publication:
United States
Language:
English

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Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ADDITIVES
AMIDES
AMINES
ANTICOAGULANTS
AUTORADIOGRAPHY
BETA DECAY RADIOISOTOPES
CARBOHYDRATES
CARBONIC ACID DERIVATIVES
CATIONS
CHARGED PARTICLES
DAYS LIVING RADIOISOTOPES
DETERGENTS
DRUGS
ELECTRON CAPTURE RADIOISOTOPES
EMULSIFIERS
HEMATOLOGIC AGENTS
HEPARIN
INTERMEDIATE MASS NUCLEI
IODINE 125
IODINE ISOTOPES
IONS
ISOTOPE APPLICATIONS
ISOTOPES
LABELLED COMPOUNDS
MEMBRANE PROTEINS
MUCOPOLYSACCHARIDES
NUCLEI
ODD-EVEN NUCLEI
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
ORGANIC SULFUR COMPOUNDS
POLYSACCHARIDES
PROTEINS
RADIOISOTOPES
RADIORECEPTOR ASSAY
RECEPTORS
SACCHARIDES
SURFACTANTS
TRACER TECHNIQUES
UREA
WETTING AGENTS