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Sircol assay kit

Manufactured by Biocolor
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Sourced in United Kingdom, Ireland

The Sircol assay kit is a colorimetric assay used to quantify the total amount of soluble collagen in biological samples. The kit utilizes a dye-binding method to measure the concentration of collagen in the sample. The assay is designed to provide a rapid and accurate measurement of collagen content.

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Lab products found in correlation

Papain, by Merck Group (3 mentions) Pepsin, by Merck Group (3 mentions) Blyscan assay kit, by Biocolor (2 mentions) Mouse tgf β1 duoset elisa, by R&D Systems (2 mentions) Quant it picogreen dsdna assay kit, by Thermo Fisher Scientific (1 mentions) Spectramax m2, by Molecular Devices (1 mentions) Sircol soluble collagen assay, by Biocolor (1 mentions) Blyscan glycosaminoglycan assay kit, by Biocolor (1 mentions) Bca assay kit, by Thermo Fisher Scientific (1 mentions) Victor 3v, by PerkinElmer (1 mentions) Lobind, by Eppendorf (1 mentions) Tgf β1, by R&D Systems (1 mentions) Genios, by Tecan (1 mentions)

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Sircol assay (47 citations) Sircol (21 citations) Sircol kit (13 citations)

16 protocols using sircol assay kit

1

Quantification of Collagen, GAG, and DNA in Cell Cultures

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For Col quantification on day 14, samples from cell groups were digested in 0.1 mg/mL pepsin (Sigma-Aldrich) in 0.2 M acetic acid at 37 °C for 16 hours. A Sircol assay kit (Biocolor Life Science Assays) was then used to quantify Col. For GAG and DNA quantification on day 14, other samples from cell groupings were digested in 125 μg/ml papain (Sigma-Aldrich) in 100 mM sodium phosphate buffer (pH 6.5) containing 10 mM L-cysteine and 10 mM EDTA at 65 °C for 16 hours. A Blyscan assay kit (Biocolor Life Science Assays) was used for GAG, and the Quant-iT™ PicoGreen™ dsDNA Assay Kit for DNA. The three kits were used following manufacturer directions as has been done in similar studies by others (Cai et al., 2015 (link); Coppola et al., 2018 (link); A. Nazempour et al., 2017 (link)).
Abusharkh H.A., Mallah A.H., Amr M.M., Mendenhall J., Gozen B.A., Tingstad E.M., Abu-Lail N.I, & Van Wie B.J. (2021). Enhanced Matrix Production by Cocultivated Human Stem Cells and Chondrocytes Under Concurrent Mechanical Strain: Coculture and Strain for Cartilage Formation. In vitro cellular & developmental biology. Animal, 57(6), 631-640.
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2

Quantification of Collagen, GAGs, and DNA

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For total collagen quantification, samples were first digested in 0.1 mg/mL pepsin (0.1% pepsin (Sigma-Aldrich) in 0.5 M acetic acid) at 60 °C for 1 h, followed by incubation at 37 °C for 15 h. Total collagen in the digest was then quantified using the Sircol assay kit (Biocolor Life Science Assays), following the manufacturer’s directions. For sulfated glycosaminoglycans and DNA quantification, samples were digested in 125 μg/mL papain (Sigma-Aldrich) in 100 mM pH 6.5 sodium phosphate buffer containing 10 mM L-cysteine and 10 mM EDTA at 60 °C for 16 h. The Blyscan assay kit (Biocolor Life Science Assays) was used to quantify glycosaminoglycans and the Quant-iT™ PicoGreen™ dsDNA Assay Kit (ThermoFisher) was used to quantify DNA, following the manufacturers’ directions.
Abusharkh H.A., Reynolds O.M., Mendenhall J., Gozen B.A., Tingstad E., Idone V., Abu-Lail N.I, & Van Wie B.J. (2021). Combining stretching and gallic acid to decrease inflammation indices and promote extracellular matrix production in osteoarthritic human articular chondrocytes. Experimental cell research, 408(2), 112841.
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3

Collagen Content Determination in Granulation Tissue

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The total collagen content in the enzymatically digested granulation tissue collected from the experimental rats was determined using a Sircol™ assay kit (Biocolor, Carrickfergus, UK).41 The assay was based on the specific binding of the dye to collagen. This methodology gives only the collagen content, free from interference of noncollagenous protein (albumin and elastin).
Bhaskar Rao A., Prasad E., Deepthi S.S., Haritha V., Ramakrishna S., Madhusudan K., Surekha M.V, & Venkata Rao Y.S. (2015). Wound healing: a new perspective on glucosylated tetrahydrocurcumin. Drug Design, Development and Therapy, 9, 3579-3588.
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4

Collagen Synthesis in Tendon Cells

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Tendon derived cells were seeded on uncompacted collagen gel samples, ELAC sutures samples and ELAC sutures samples with PDGF-BB. Samples were cultured in ascorbic acid free media for 21 days. Every 3 days the media was collected and fresh media was replaced. All collected media for each group from different time points were mixed and total synthesized collagen by cells over 21 days was measured using the Sircol assay kit for measuring the amount of collagen in the culture medium (Sircol Soluble Collagen Assay, Biocolor Life Science Assays). A micro-plate reader (Spectramax M2, Molecular Devices) was used to measure the absorbance of the samples at 555 nm wavelength. Total collagen amount synthesized by cells were calculated by using the calibration curve plotted based on standard samples absorbance. Tests were repeated in triplets.
Younesi M., Donmez B.O., Islam A, & Akkus O. (2016). Heparinized Collagen Sutures for Sustained Delivery of PDGF-BB: Delivery Profile and Effects on Tendon-derived Cells In-Vitro. Acta biomaterialia, 41, 100-109.
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5

Quantifying Collagen and TGF-β in BALF

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Total acid soluble collagen content in cell free BALF was determined using the Sircol assay kit (Biocolor, Ltd; Carrickfergus, UK) according to the manufacturer’s instructions. TGFβ concentration in the cell free BALF was calculated using Mouse TGFβ 1 DuoSet Elisa (R&D Systems Cat DY1679–05) according to the manufacturer’s instructions.
Rodriguez L.R., Tang S.Y., Barboza W.R., Murthy A., Tomer Y., Cai T.Q., Iyer S., Chavez K., Das U.S., Ghosh S., Dimopoulos T., Babu A., Connelly C., FitzGerald G.A, & Beers M.F. (2023). Disruption of Prostaglandin F2α Receptor Signaling Attenuates Fibrotic Remodeling and Alters Fibroblast Population Dynamics in A Preclinical Murine Model of Idiopathic Pulmonary Fibrosis. bioRxiv.
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6

Determining Collagen Content in Lung Tissue

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The Sircol assay kit (Biocolor Ltd., Belfast, Northern Ireland, UK) was utilized to assess collagen content of lung tissue. Briefly, the specimens previously frozen in liquid nitrogen and stored at -80°C were thawed, and assays were performed as instructed by the manufacturer. Collagen concentrations were determined from a standard curve.
Lee S.J., Yi C.O., Heo R.W., Song D.H., Cho Y.J., Jeong Y.Y., Kang K.M., Roh G.S, & Lee J.D. (2015). Clarithromycin Attenuates Radiation-Induced Lung Injury in Mice. PLoS ONE, 10(6), e0131671.
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7

Glycosaminoglycan and Collagen Quantification

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Constructs harvested at 24 and 52 weeks were digested with 125 mg/mL papain (Sigma) in 50 mM phosphate buffer supplemented with 2 mM N-acetyl cysteine and 2 mM ethylenediaminetetraacetic acid (EDTA) at 65°C overnight. GAGs was assessed by the Blyscan Glycosaminoglycan Assay Kit (Biocolor) following the manufacturer's instructions; absorbance was measured at 656 nm using a microplate reader. Total collagen content was determined using the Sircol Assay Kit (Biocolor) according to the manufacturer's suggested protocol. Briefly, the samples were digested with 0.1 mg/mL pepsin (Sigma) supplemented with 0.5 M acetic acid (to keep enzyme activity) at 4°C overnight; bovine collagen-I solution (Biocolor) was used as a standard; absorbance was measured at 555 nm.
Zhang W., Lian Q., Li D., Wang K., Hao D., Bian W., He J, & Jin Z. (2014). Cartilage Repair and Subchondral Bone Migration Using 3D Printing Osteochondral Composites: A One-Year-Period Study in Rabbit Trochlea. BioMed Research International, 2014, 746138.
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8

Fibroblast Collagen Secretion Modulation

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Confluent serum-deprived fibroblasts were stimulated with TGF-β1 (2.5 ng/ml) and incubated in the presence of either IFN-γ, PFD or both for the indicated time periods. Soluble collagen secretion (collagens type I to V) in the conditioned media was determined using a Sircol™ Assay kit (Biocolor; Carrick Fergus, UK) according to the manufacturer’s instructions.
Vu T.N., Chen X., Foda H.D., Smaldone G.C, & Hasaneen N.A. (2019). Interferon-γ enhances the antifibrotic effects of pirfenidone by attenuating IPF lung fibroblast activation and differentiation. Respiratory Research, 20, 206.
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9

Quantifying Soluble Collagen in Cell Cultures

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Total soluble collagen in the cell culture supernatant was measured using the Sircol assay kit (Biocolor, Newtownabbey, United Kingdom) following the manufacturer’s instructions. To minimize the influence of cell number, the level of soluble collagen was normalized to total protein in the cell lysate measured using a BCA assay kit (Pierce Biotechnology, Rockford, IL).
Omori K., Hattori N., Senoo T., Takayama Y., Masuda T., Nakashima T., Iwamoto H., Fujitaka K., Hamada H, & Kohno N. (2016). Inhibition of Plasminogen Activator Inhibitor-1 Attenuates Transforming Growth Factor-β-Dependent Epithelial Mesenchymal Transition and Differentiation of Fibroblasts to Myofibroblasts. PLoS ONE, 11(2), e0148969.
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10

Quantifying Collagen in Esophageal Tissues

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The amount of soluble and insoluble collagen in the lyophilized tissue sample of normal and decellularized esophagi (n = 7) tissues were measured by the Sircol Assay Kit (Biocolor, Carrickfergus, UK). Collagen was extracted by acid–pepsin extraction procedure according to the instruction given with the kit and precipitated by adding acid neutralizing reagent, followed by isolation and concentration reagent. Furthermore, the precipitate was incubated with collagen dye reagent. This complex was further recovered by the addition of alkali reagent and measured at 555 nm on a plate reader. All reagents used were provided with the kit.
Nayakawde N.B., Methe K., Banerjee D., Berg M., Premaratne G.U, & Olausson M. (2020). In Vitro Regeneration of Decellularized Pig Esophagus Using Human Amniotic Stem Cells. BioResearch Open Access, 9(1), 22-36.
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