Sircol™ Collagen Assay

Now sensitive to 1μg!

Sircol Collagen Kit FAQs

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:

in vivo

- collagen, soluble in cold acid or pepsin, recovered from mammalian soft tissues, cartilages and fluids.

in vitro

- 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).

Sircol Collagen Assay Manual Sircol Collagen Assay Citations

Sircol Method Summary

1. Test materials, standards and reagent blanks (100µl) are mixed with Sircol Dye reagent (1ml) in microcentrifuge tubes for 30 minutes. The collagen-dye complex formed precipitates out of solution.
2. The complex is packed firmly to the bottom of the tubes by centrifugation at 12,000g for 10 minutes. Unbound dye is removed by inverting and draining tubes.
3. Ice-cold acid-salt wash (750µl) is gently added to remove unbound dye from pellet surface and microcentrifuge tube. The tubes are centrifuged at 12,000rpm for 10 minutes and the wash drained off.
4. Bound dye is released and dissolved via addition of alkali reagent (250µl or 1000µl depending on pellet size) and vortex mixing for 10 minutes.
5. Released dye is measured spectrophotometrically at 555nm or colorimetrically with a blue-green filter using a multiwell plate reader.

Collagen Standards: 0, 5, 10, 15 μg

Standards after adding 250μl of Sircol Alkali Reagent

Tube aliquots (200μl) transferred to 96-well microplate

Sircol Kit Components

• Dye Reagent: Sirius Red in picric acid
• Dye release agent: alkaline reagent containing 0.5M sodium hydroxide
• Reference Standard: acid soluble Type 1 collagen in 0.5M acetic acid. Concentration: 500 µg/ml
• Acid-Salt Wash Reagent (Concentrate): contains acetic acid, sodium chloride and surfactants
• Acid Neutralising Reagent: contains TRIS-HCl and NaOH.
• Collagen Isolation & Concentration Reagent: contains polyethylene glycol in a TRIS-HCl buffer, pH 7.6

Sircol Collagen Products

Standard Assay Kit (120 assays) Product Code: S1000

Economy Pack (475 assays) Product Code: S5000

Sircol Dye Reagent (3 x 120ml) Product Code: S1005

Sircol Dye Release Reagent (3 x 150ml) Product Code: S1007

Sircol Bovine Collagen Reference Standard (0.5mg/ml, 3 x 5ml)

Product Code: S1010

Sircol Rat Collagen Reference Standard (0.5mg/ml, 3 x 5ml)

Product Code: S1011

Sircol Collagen Assay (120 assays) (Rat Collagen) Product Code: S1111

Sircol Dye & Dye Release Reagents (2 x 2 x 120ml) Product Code: S1500

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Sircol Collagen Kit FAQs