Biocolor Sircol™ Insoluble Collagen Assay - Frequently Asked Questions (FAQ)

Here you will find answers to many of the frequently asked questions about Biocolor Sircol™ Insoluble Collagen Assay.

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What is the Sircol™ Insoluble Collagen Assay?

The Biocolor Sircol™ INSOLUBLE Collagen Assay Kit is a dye-binding assay designed for accurate quantification and measurement of insoluble or cross-linked collagen fibers. It is ideal for analyzing materials from sources such as tissues, bone, and calcified tissue.

General Questions

What absorbance values can samples be measured at?

Absorbance spectrum maximum for Sircol dye in alkali reagent is 556 nm; however, absorbance settings within the range of 520-570 nm will give suitable readings.

Why does the Sircol Insoluble Collagen Assay demonstrate lower absorbance values when compared to the Sircol Soluble Collagen Assay?

Maximal binding of the Sircol dye occurs in collagens possessing intact triple helix organization since the highly ordered [Gly-X-Y]n helical structure of tropocollagen contributes to dye binding. Affinity is progressively reduced during heat denaturation (> 45 ºC) due to the unwinding of the triple helix and formation of random chains. Since samples and standards for the Sircol Insoluble Collagen Assay are in a denatured state (necessary to convert insoluble "solid" samples into the liquid phase for assay) they will therefore bind less dye than the same amount of material in the non-denatured state.

The collagen pellet is much smaller than the pellets in the S1000 Sircol Soluble Collagen Assay, why is this?

Samples in the Sircol Insoluble Collagen Assay are gently denatured via treatment with Sircol Fragmentation Reagent. The denaturation causes some loss of collagen structure. The dye-bound pellet will therefore be more "dense" as it is able to pack more tightly following centrifugation.