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Soluble Collagen Assay Kit (ab241015)

  • Datasheet
  • SDS
  • Protocol Booklet
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Collagen I Standard curve.
  • Estimation of acid-soluble collagen content in rat tissues.
  • Estimation of secreted soluble collagen in cultured cell growth medium (DMEM/F12 medium).

Key features and details

  • Detection method: Fluorescent

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Overview

  • Product name

    Soluble Collagen Assay Kit
    See all Collagen kits
  • Detection method

    Fluorescent
  • Product overview

    Soluble Collagen Assay Kit (ab241015) detects newly synthesized (acid-soluble) collagen levels in various biological samples and cultured cell medium.


    The assay involves extraction of soluble collagen in acetic acid, followed by enzymatic degradation of collagen into glycine-rich oligopeptides, which are quantified using a fluorogenic reagent and developer solution that selectively react with the N-terminal glycine fragments to form a stable fluorescent complex (Ex/Em = 376/468 nm).


    The assay is more sensitive and selective than dye-binding (Picrosirius Red) soluble collagen assays, is simple to perform and has a linear range from 0.05 – 2 μg collagen per well (2.5 μg/mL to 100 μg/mL in a 20 μL sample volume).

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components 100 tests
    Collagen Assay Buffer 1 x 25ml
    Collagen I Standard (2 mg/ml) 1 x 100µl
    Collagenase Enzyme Mix 1 vial
    Detection Reaction Buffer 1 x 10ml
    Developer Solution 1 x 500µl
    Peptide Labelling Reagent 1 vial

Associated products

    Images

    • Collagen I Standard curve.
      Collagen I Standard curve.
      Collagen I Standard curve.
    • Estimation of acid-soluble collagen content in rat tissues.
      Estimation of acid-soluble collagen content in rat tissues.

      Rat leg muscle, lung and heart samples were homogenized in 0.5 M acetic acid, incubated overnight at 4C to solubilize collagen and neutralized with 0.5 M NaOH. Collagen levels (calculated as µg collagen/mg wet tissue) for the samples were: 3.83 ± 0.47 µg/mg for muscle, 1.47 ± 0.22 µg/mg for lung and 2.85 ± 0.08 µg/mg for heart.

    • Estimation of secreted soluble collagen in cultured cell growth medium (DMEM/F12 medium).
      Estimation of secreted soluble collagen in cultured cell growth medium (DMEM/F12 medium).

      3T3-L1 fibroblasts were grown to ~80% confluence in T-75 flasks and then treated with either vehicle or TGF-β (100 ng/ml), a cytokine known to stimulate synthesis and secretion of tropocollagen fibrils. After 48 hours of treatment, the culture medium was removed, centrifuged to remove debris and was assayed directly (each 10 µl clarified medium, undiluted). TGF-β treatment resulted in a roughly 1.5- fold increase in collagen secretion. Data are mean ± SEM of 3 replicates, assayed according to the kit protocol.

    Protocols

    • Protocol Booklet

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet
    • SDS
  • References (0)

    Publishing research using ab241015? Please let us know so that we can cite the reference in this datasheet.

    ab241015 has not yet been referenced specifically in any publications.

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