Products ❯ Fibrosis ❯ QuickZyme Human MMP-2 Activity Assay Kit

QuickZyme Human MMP-2 Activity Assay Kit

$0.00

  • Catalog #: QZBmmp2Hv2
  • Method: Chromogenic Assay
  • Packaging: Kit/96 tests
  • Type: Kit
  • Use: Research Use Only (RUO)

MMP-2 (also known as 72 kDa type IV collagenase, Gelatinase A) has a broad range of substrate specificity for denatured collagens (gelatins) and native collagens (types IV, V, VII and X). Human MMP-2 has a MW of 72 kDa (proform) and 62 kDa (active form). MMP-2 is produced by a variety of cell types including fibroblasts, chondrocytes, endothelial and epithelial cells. Studies support the use of MMP-2 and MMP-9 as biomarkers for cancer, liver fibrosis, pulmonary hypertension, and preeclampsia, among others.

The QuickZyme human MMP-2 immunoenzymatic assay enables you to specifically measure in biological samples both active MMP-2, as well as (pro)MMP-2, which is activated on the plate by APMA. It can be used for the measurement of MMP-2 activity in various biological samples, such as conditioned culture media, tissue homogenates, serum, plasma and urine.

This product ships on dry ice.

Assay Principle

Active MMP-2
Measurement of active human MMP-2 Standards, controls and biological samples are pipetted into the pre-coated plate. Human MMP-2 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-2 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-2
Measurement of total MMP-2 is done similarly to the measurement of active MMP-2. After binding of MMP-2 to the antibody-coated plate, bound MMP-2 is first activated by adding APMA, resulting in the activation of pro-MMP-2. The activity of total MMP-2 (the newly activated MMP-2 and the already active MMP-2 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.

96 well microwell plate 12×8 well ready-to-use strips coated with antiMMP-2
Assay buffer 30 ml bottle contains 20 ml ready-to-use Tris-HCl buffer
Standard tube contains 50 µl of 640 ng/ml pro-MMP-2 (human)
p-Aminophenylmercuric acetate (APMA) tube contains 17.5 mg APMA, see safety data sheet supplied
Detection enzyme tube contains 600µl detection enzyme in Tris-HCl buffer
Substrate tube contains 1000 µl peptide substrate in demineralized water
Wash buffer 30 ml bottle contains 20 ml 25x concentrated phosphate buffer

Note that the protocol requires the use of Dimethyl Sulphoxide (DMSO).

Materials required but not supplied

  • Single and/or multichannel pipettes with disposable polypropylene tips.
  • Polypropylene tubes (Eppendorf tubes).
  • Glass measuring cylinder 500 ml.
  • Distilled or demineralized water.
  • Microplate shaker.
  • Refrigerator at 2-8 °C.
  • Dimethyl Sulphoxide (DMSO).
  • (Microtitre plate) incubator at 37°C.
  • Automatic plate washer or wash bottle (optional).
  • Microplate reader capable of measuring at 405 nm
  • Measures endogenous active MMP-2 (naturally occurring) or total active MMP-2 (following activation with APMA).
  • Samples: cell culture conditioned medium, serum, plasma, urine and tissue homogenates.
  • Quantitative.
  • Range: 0.02 – 16 ng/ml.
  • Sensitivity: 0.04 ng/ml for 2 h incubation with detection reagent;  0.02 ng/ml for 6 h incubation with detection reagent; 4 pg/ml for o/n incubation
  • Ease-of-use: Equivalent to ELISA.
  • Overnight protocol.
  • Store kit at -20°C, store standard at -70°C

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