Soluble Collagen Assay Sircol™
Sircol™ Soluble Collagen Assay
The Sircol™ Collagen Assay is a dye-binding method for the analysis of acid and pepsin-soluble collagens. The Sircol Assay is suitable for monitoring collagen produced in situ or during in-vitro cell culture and in-vitro extracellular matrix, (ECM), formation:
– collagen, soluble in cold acid or pepsin, recovered from mammalian soft tissues, cartilages and fluids.
– collagen, soluble in cold acid or pepsin, released into cell culture medium during cell growth and cell maintenance.
– collagen, soluble in cold acid or pepsin, recovered from newly formed extracellular matrix that has been deposited onto cell culture treated plastic surfaces, (T-flasks and microwell plates).
The assay can assess the rate of newly synthesised collagen produced during periods of rapid growth and developement. New collagen is also generated during inflammation, wound healing and tumour developement.
(a) Sets of collagen standards, Low range 0, 5, 10 & 15μg and High range 0, 15, 30 & 45μg, following collagen-dye mixing, centrifuging and removal of unbound dye, (duplicates not shown).
(b) Low Standards after adding 250μl Alkali Reagent. High standards after adding 1000μl Alkali Reagent.
(c) Tube aliquots (200μl) transferred to 96 well microplate.
Q. What is the sensitivity of the Sircol assay?
A. 1.0µg collagen. Quantities as low as 1.0µg of collagen can be recovered from a 1 ml volume if the Collagen Isolation & Concentration protocol is used (See Sircol assay manual for more detail).
Q. How long does the Sircol assay take to run?
A. 1.5 hours
Q. What forms of collagen does the Sircol assay measure?
A. Acid-soluble and pepsin-soluble collagens.
Q. What types of collagen can the Sircol assay be used with?
A. Mammalian collagens, Types I to V.
Q. What sort of collagen standard should I use?
A. The Kit containing bovine collagen is suitable for use with all mammalian cells.
Q. What absorbance values can samples be measured at?
A. Absorbance spectrum maximum for Sircol dye in alkali reagent is 555 nm however absorbance settings within the range 520-570 nm will give suitable readings.
Q. How do I prepare samples for measurement with the Sircol assay?
A. Preparation details for cell culture and tissue samples can be found in the assay manual.