MMP-9 (also known as type neutrophil gelatinase, IV collagenase, Gelatinase B) has a broad range of substrate specificity for denatured collagens (gelatins), native collagens (types IV, V and XI), as well as elastin. Human MMP-9 has a MW of 92 kDa (pro-form) and 82 kDa (active form). MMP-9 is produced by a variety of cell types including monocytes, macrophages, fibroblasts, neutrophils, osteoclasts, chondrocytes, keratinocytes, 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 mouse MMP-9 activity assay enables you to specifically measure in biological samples both active mouse MMP-9, as well as (pro)MMP-9 which is activated on the plate by APMA. It can be used for the measurement of MMP-9 activity in various biological samples, such as conditioned culture media, tissue homogenates, serum, plasma and urine.
This product ships on dry ice.
Active MMP-9 Measurement of active mouse MMP-9 Standards, controls and biological samples are pipetted into the pre-coated plate. Mouse MMP-9 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-9 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 mouse MMP-9 Measurement of total MMP-9 is done similarly to the measurement of active MMP-9. After binding of MMP-9 to the antibody-coated plate, bound MMP-9 is first activated by adding APMA, resulting in the activation of pro-MMP-9. The activity of total MMP-9 (the newly activated MMP-9 and the already active MMP-9 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 kit contains:
- 96 well microwell plate – 12×8 well ready-to-use strips coated with anti-MMP-9
- Assay buffer – 125 ml bottle contains 100 ml ready-to-use TrisHCl buffer
- Standard – tube contains 20 µl of approximately 500 ng/ml proMMP-9 (mouse), exact concentration can be found on Standard Lot Specification Sheet.
- 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
The following materials and equipment are 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-9 (naturally occurring) or total active MMP-9 (following activation with APMA).
- Samples: cell culture conditioned medium, serum, plasma, urine and tissue homogenates.
- Range: 4 – 4000 pg/ml.
- Sensitivity: 20 pg/ml for 1 h incubation with detection reagent; 1 pg/ml for 6 h incubation with detection reagent.
- Ease-of-use: Equivalent to ELISA.
- Overnight protocol.
- Store at -15 to -30°C