1 simple way to rapidly quantify Collagen: Sircol Collagen Assay

Collagen is a structural protein expressed in mammalian tissues (ex. skin, muscles, tendons, ligaments, cartilages, blood vessels…).  Together with Elastin and Keratin, Collagen fibres give connective properties to mammalial tissues ensuring their physiological form but also firmness, strength and flexibilityNeedless to say, Collagen is trendy… a simple search in Google using the term “Collagen” clearly shows the economical dimension of this protein in well-being.

However, far beyond this, Collagen is a key element in pharmaceutical and cosmetology R&D projects. The research projects focused on Collagen show its tremendous impacts on tissue regeneration, skin repair, wound healing, allergy and inflammation, surgery, ageing diseases…

Collagen Type II (IHC-P staining)

Collagen Type II staining in FFPE human bronchiolar cartilage. Source: GeneTex.

Monitoring Collagen during experimental approaches is crucial. However, Collagen assays are indeed quite challenging:

  • Various types of Collagens are identified  (ex. Collagen type I in skin, tendon, bone…, Collagen type II in cartilages, Collagen Type III in reticular fibers with Type I, Collagen type IV in basal lamina…, Collagen Type V in cell surfaces, hair, placenta…)
  • Collagen’s chemistry and structure make it not soluble and difficult to purify
  • Current quantification assays are long and tedious

To overcome these hurdles, several protocols to quantify mammalian Collagens (Types I to V) by measuring Acid-Soluble Collagen (ASC) and Pepsin-Soluble Collagen (PSC) forms have been designed. The most robust Collagen assay is the Sircol Assay, for 3 reasons:

#1 – Sircol Collagen Assay detects up to 1.0 µg of Collagen (Type I to V) in only 1.5 hours

The Sircol Assay is a dye-binding method designed for the analysis of ASC and PSC by absorbance readings within the range 520-570 nm.

This Collagen Assay is ideal for in vivo and in vitro Collagen monitoring:

  • in vivo for Collagen recovered from mammalian soft tissues, cartilages and fluids…
  • in vitro for Collagen released into cell culture medium during cell growth, cell maintenance…
  • in vitro for Collagen recovered from newly formed extracellular matrix that has been deposited onto cell culture treated plastic surfaces…

Quantities as low as 1.0µg of collagen can be recovered from a 1 ml volume with a dedicated Collagen Isolation & Concentration protocol.

#2 –  Sircol Collagen Assay is well established and recognised within the Scientific community

There are dozens of scientific publications using the Sircol Assay to measure Collagen in numerous Human, Rat and Mouse samples:

Tissues: Aortic Dermal, Bladder, Cardiac, Intervertebral disc, Kidney, Liver, Lung, Wound tissue…

Cells and cell lines:  Adipose Stem Cells, Alveolar Aortic Endothelial Cells, Bone Marrow Stromal Cells, Breast Cancer Cells, Chondrocyte Induced Bone Marrow Stromal Cells, Dental Pulp Cell,s Dermal Fibroblasts, Epithelial Cells, Crohn’s Disease Fibroblasts, Myofibroblasts , Osteoblasts and Osteoclasts,  Periodontal Ligament Cells, Skin Fibroblasts, Tenocytes, Umbilical Cord Mesenchymal Stromal cells…

#3 Collagen measurements with Sircol assays can be outsourced to CRO

Another option is to outsource Collagen quantifications of samples to an ISO 9001:2008 Certified Contract Research Organization (CRO). CRO laboratories with extended R&D competencies in proteomics, genomics and cell culture and are fully able to perform such quantification assays. For example, tebu-bio’s laboratory lab staff regularly perform assays related to Collagen measurement with the Sircol protocol, but also tests related to anti-ageing, anti-oxidant and anti-cellulite tests (oxidative markers, MMPs, ORAC, ABTS, PPAR, ECM…).

Want to rapidly and simply quantify Collagen with the Sircol Assay?

Quantify Collagen levels in your samples by yourself, or by outsourcing it to tebu-bio’s lab staff who are experienced in running the Sircol Assay.

Colorimetric protein Assay plate

Collagen Sircol assays and other biomarker Assays outsourced to tebu-bio laboratories are eligible to tax relief on expenses for R&D under the French Research Tax credit System (CIR).

Whatever your choice, it’s as easy as ABC and… robust. So to adopt Collagen quantification with the Sircol Assay!

 

Trackbacks

  1. […] assay using Sirius Red dye (Sircol dye) in a convenient manner (we explored this method in a recent post).  Nevertheless, up to now it has been a challenge to measure insoluble collagen. During periods […]

  2. […] the effects of compounds on molecular pathways and for quantifying skin-related biomarkers (Collagen assays, Elastin measurements, Hyaluronic acid quantifications…) have been made available. To further […]

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