Collagen is the most abundant protein in mammals, accounting for nearly 30% of body protein content. Collagen is an extracellular scaffolding protein found primarily in skin, tendon, bone, cartilage, ligament and muscle. There are over 20 characterized collagen types, but Types I, II and III comprise more than 90% of mammalian collagen. Collagen exists as a triple helix, with a repeating Gly-X-Hyp tripeptide sequence, where X is any amino acid and Hyp is hydroxyproline. Hydroxyproline is found almost entirely in collagen and is thus a direct measure of the amount of collagen present in samples. Multiple diseases affect collagen turnover, including tumor invasion and metastasis, rheumatoid arthritis, cardiopulmonary fibrosis and muscular dystrophy. AkrivisBio’s Total Collagen Assay is a convenient method for quantifying small amounts of collagen from a variety of biological samples. In the assay, samples are hydrolyzed to yield free hydroxyproline, which is oxidized and reacted to give a brightly-colored chromophore with an absorbance peak at 560 nm. Our assay employs a proprietary oxidizing acid reagent which avoids the use of perchloric acid. The assay detects down to 0.5 µg collagen per well in a 96-well format.
