MMP-7 (also known as matrilysin, matrilysin 1 or pump 1) has a broad range of substrate specificity including collagen type IV, elastin, fibronectin, laminin, nidogen, tenascin, osteonectin, MBP, decorin and versican. Human MMP-7 has a MW of 29 kDa (pro-form) and 19 kDa (active form). MMP-7 is produced by a variety of cell types including osteoclasts, keratinocytes epithelial cells and macrophages. Studies also support MMP-7 as a biomarker in the lung for idiopathic pulmonary fibrosis and interstitial lung disease, in the kidney as a urinary marker for acute kidney injury and kidney fibrosis, as well as in the liver as a marker related to liver fibrosis and NAFLD. Furthermore, serum MMP-7 activity is emerging as a prime candidate biomarker for the liver disease, biliary atresia.
The QuickZyme human MMP-7 activity assay enables you to specifically measure in biological samples both active MMP-7, as well as (pro)MMP-7, which is activated on the plate by APMA. It can be used for the measurement of MMP-7 activity in various biological samples, such as conditioned culture media, tissue homogenates, serum, plasma and urine.
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Active MMP-7 Measurement of active human MMP-7 Standards, controls and biological samples are pipetted into the pre-coated plate. Human MMP-7 present in the biological sample is captured by the antibody. After washing, the pro-detection enzyme is added. This is activated by the active MMP-7 into an active detection enzyme. The active detection enzyme is able to cleave the chromogenic substrate, resulting in generation of a yellow color that can be measured at 405 nm using an ELISA plate reader.
Total human MMP-7 Measurement of total MMP-7 is done similarly to the measurement of active MMP-7. After binding of MMP-7 to the antibody-coated plate, bound MMP-7 is first activated by adding APMA, resulting in the activation of pro-MMP-7. The activity of total MMP-7 (the newly activated MMP-7 and the already active MMP-7 present in the sample) is measured by adding the detection enzyme, followed by the addition of chromogenic substrate. The released color can be measured at 405 nm using an ELISA plate reader.
Each 96 well assay contains:
- 96 well microwell plate – 12×8 well ready-to-use strips coated with Rabbit-anti-Goat IgG (RAG)
- Antibody stock – anti-MMP-7 stock solution, dilute 100x before use
- Assay buffer – 125 ml bottle contains 100 ml ready-to-use TrisHCl buffer
- Standard – tube contains 50 µl of 640 ng/ml pro-MMP-7 (human)
- p-Aminophenylmercuric acetate (APMA) – tube contains 17.5 mg APMA, see safety data sheet supplied
- Detection enzyme – tube contains 600 µl detection enzyme in Tris-HCl buffer
- Substrate – tube contains 1000 µl peptide substrate in demineralized water
- Wash buffer – 30 ml bottle contains 25 ml 20x concentrated phosphate buffer
- Measures endogenous active MMP-7 (naturally occurring) or total active MMP-7 (following activation with APMA).
- Samples: conditioned culture medium, serum, urine and tissue homogenates.
- Range: 0.004 – 16 ng/ml.
- Sensitivity: 30 pg/ml for 3 h incubation with detection reagent; 4 pg/ml for 25 h incubation with detection reagent.
- Ease-of-use: Equivalent to ELISA.
- Overnight protocol.
- Store kit at -20°C, store standard at -70°C