Collagen Degradation/Zymography Assay Kit (Fluorometric) (ab234624)
Key features and details
- Assay type: Quantitative
- Detection method: Fluorescent
- Platform: Microplate reader
- Sample type: Cell Lysate, Tissue Homogenate, Tissue Lysate
Overview
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Product name
Collagen Degradation/Zymography Assay Kit (Fluorometric) -
Detection method
Fluorescent -
Sample type
Cell Lysate, Tissue Homogenate, Tissue Lysate -
Assay type
Quantitative -
Product overview
Collagenase (Collagen Degradation/Zymography) Assay Kit (Fluorometric) (ab234624) employs a highly quenched collagen substrate which upon cleavage by a suitable collagenase releases a fluorophore, which can be easily quantified using a conventional microplate reader. This method is substrate-specific, simple, fast, high-throughput adaptable and amenable to the sensitive detection of collagenase activity (as low as 0.6 mCDU for bacterial collagenase) in biological samples.
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Notes
Collagenases, members of the matrix metalloproteinase (MMP) family, are endopeptidases that digest native collagen in the triple helix region and are involved in the physiological and pathological turnover of connective tissues. Collagens are the major fibrous component of animal extracellular connective tissue. Bacterial collagenases differ from vertebrate collagenases in that they exhibit broader substrate specificity. Collagenases may play a role in facilitating tumor cell invasion of the extracellular matrix at multiple stages of the metastatic process. Collagenases have also been approved for medical uses for the treatment of Dupuytren's contracture, Peyronie's disease and wound healing.
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Platform
Microplate reader
Properties
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Storage instructions
Store at -20°C. Please refer to protocols. -
Components 100 tests Cell Lysis Buffer 1 x 25ml Collagenase Assay Buffer 1 x 25ml Collagenase Substrate 1 vial Enzyme Positive Control 1 x 10µl FITC Standard (5 mM) 1 x 10µl -
Research areas
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Relevance
Function: Cleaves collagens of types I, II, and III at one site in the helical domain. Also cleaves collagens of types VII and X. In case of HIV infection, interacts and cleaves the secreted viral Tat protein, leading to a decrease in neuronal Tat's mediated neurotoxicity. Domain: There are two distinct domains in this protein; the catalytic N-terminal, and the C-terminal which is involved in substrate specificity and in binding TIMP (tissue inhibitor of metalloproteinases). The conserved cysteine present in the cysteine-switch motif binds the catalytic zinc ion, thus inhibiting the enzyme. The dissociation of the cysteine from the zinc ion upon the activation-peptide release activates the enzyme. PTM: Undergoes autolytic cleavage to two major forms (22 kDa and 27 kDa). A minor form (25 kDa) is the glycosylated form of the 22 kDa form. The 27 kDa form has no activity while the 22/25 kDa form can act as activator for collagenase. Similarity: Belongs to the peptidase M10A family. Contains 4 hemopexin-like domains. -
Cellular localization
Secreted, extracellular space, extracellular matrix -
Alternative names
- CLG
- Fibroblast collagenase
- matrix metallopeptidase 1
see all
Images
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Example Standard Curve.
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Collagenase activity with different amounts of Enzyme Positive Control. -
Collagenase activity in rat kidney, liver lysates along with Hela and U937 cell lysates. n=3.
