MMP-14, also known as MT- MMP or MT1-MMP, is a membrane type MMP and contains a transmembrane domain. MMP-14 has a role in the activation of the proforms of MMP-2 and MMP-13. Furthermore, MMP-14 is able to directly degrade certain matrix proteins such as laminin, fibronectin, vitronectin and interstitial collagens. Human MMP-14 consists of 562 amino acid residues including a propeptide of 91 residues. and is most likely intracellularly activated by furin-like enzymes and transported to the membrane as an active enzyme. MMP-14 is thought to be released from the cell membrane by proteolytic cleavage. The QuickZyme human MMP-14 activity assay enables you to specifically measure active human MMP-14. It can be used for the measurement of MMP-14 activity in various biological samples, such as cultured cells and tissue homogenates.
MMP-14 is believed to be involved in bone and tooth development, tumor cell invasion and cell migration and in shedding of a variety of cell surface proteins. Recent work points to an involvement of MMP-14 in plaque vulnerability. A variety of studies show MMP-14 as a biomarker for metastasis and fibrosis in the liver.
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Active MMP-14 Measurement of active human MMP-14 Standards, controls and biological samples are pipetted into the pre-coated plate. Human MMP-14 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-14 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.
Each 96 well kit contains:
- 96 well microwell plate – 12×8 well non-coated
- Blocking buffer – 15 ml ready to use
- Antibody stock solution- 20 µl 5.9 mg/ml
- Assay buffer – 30 ml bottle contains 20 ml ready-to-use Tris-HCl buffer
- Extraction buffer – 125 ml bottle contains 100 ml ready-to-use Tris-HCl buffer with Triton-X-100
- Standard – tube contains 50 µl of 640 ng/ml MMP-14 (human); store at -70°C
- Detection enzyme – tube contains 600 µl detection enzyme in Tris-HCl buffer; store at -20 °C
- Substrate – tube contains 1000 µl peptide substrate in demineralized water
- Wash buffer – 30 ml bottle contains 20 ml 25x 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.
- Freezer -20 °C and -70 °C
- (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-14 / MT1-MMP (naturally occurring) in complex biological samples.
- Samples: cell extracts, tissue homogenates, tissue extracts.
- Range: 0.1 to 16 ng/ml
- Sensitivity: 0.5 ng/ml (2 h incubation with detection reagent); 0.1 ng/ml (5 h incubation with detection reagent).
- Ease-of-use: Equivalent to ELISA.
- Overnight protocol.
- Store kit at -20°C, store standard at -70°C