An assay for heparin by decrease in color yield (DECOY) of a protein-dye-binding reaction.
The interference by heparin and some related molecules with the well-known Bradford dye-binding assay for proteins is used as the basis of a rapid, sensitive method for the quantitation of these polysaccharides. Whereas the available methods for the assay of glycosaminoglycans have lacked specificity for sulfated polyanions in general and for heparin in particular, the procedure described here distinguishes among different uronic acid derivatives and, when performed in conjunction with heparinase digestion or cetylpyridinium chloride precipitation, can be used to determine heparin content of complex biological samples.
Khan
M Y
MY
Department of Cell Biology and Anatomy, New York Medical College, Valhalla 10595.
Newman
S A
SA
D000818:Animals; D002417:Cattle; D002594:Cetylpyridinium; D055598:Chemical Phenomena; D002621:Chemistry; D003124:Colorimetry; D004396:Coloring Agents; D005966:Glucuronidase; D006026:Glycoside Hydrolases; D006493:Heparin / Q000032:analysis*; D008722:Methods; D008970:Molecular Weight; D011485:Protein Binding; D015203:Reproducibility of Results; D012394:Rosaniline Dyes; D014574:Uronic Acids; D000099175:Heparanase
D002594:Cetylpyridinium; D004396:Coloring Agents; D005966:Glucuronidase; D006026:Glycoside Hydrolases; D006493:Heparin; D012394:Rosaniline Dyes; D014574:Uronic Acids; D000099175:Heparanase; C004692:coomassie Brilliant Blue