Description
Key facts
Detection method:Fluorescent,
Sample types:Purified protein, Cell culture supernatant,
Assay time:60m,
Assay Platform:Microplate reader
Product details:
How the assay works
The MMP assay protocol uses a fluorescence resonance energy transfer (FRET) peptide as a generic MMP activity indicator. In the intact FRET peptide, the fluorescence of one part is quenched by another. After cleavage into two separate fragments by MMPs, the fluorescence is recovered.
With excellent fluorescence quantum yield and longer wavelength, the probe is much more sensitive than an EDANS/Dabcyl FRET substrate. The probe signal can be easily read by a fluorescence microplate reader at Ex/Em = 490/525 nm.The kit measures total MMP activity. It does not give an individual read-out for each MMP. The substrate peptide is a sequence that is recognized by all MMPs. A specific inihibitor would need to be included to determine the activity of a specific MMP enzyme.
MMP Activity Assay Kit protocol summary
- Add appropriate controls or test samples (50 µL)
- Pre-incubate for 10 - 15 minutes
- Add MMP Green™ substrate working solution (50 µL)
- Skip incubation step for kinetic reading or incubate for 30 to 60 minutes for end point reading
- Monitor fluorescence intensity at Ex/Em = 490/525 nm
How other researchers are using
MMP Activity Assay Kit has been used in a variety of sample type including:
- SK-OV-3 and ES-2 cells 1
- Human tendon cells, 2
- Human Glioma Cell Culture 3
References:
1 - Wu C t al. 2023; 2 - Mousavizadeh R et al. 2023; 3 - Jung J et al. 2024.
Related and recommended products
See other alternative MMP activity assay kits:
- MMP Activity Assay Kit (Fluorometric - Red)
ab112147
- MMP-3 Activity Assay Kit (Fluorometric)
ab118972
REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions--20°C, Appropriate long-term storage conditions--20°C, Storage information--20°C
Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
Matrix Metalloproteinases (MMPs) are a group of enzymes that break down extracellular matrix components. These are sometimes called matrixins. MMPs have a mass typically ranging from 19 to 120 kDa depending on the specific MMP enzyme. They are expressed in various tissues throughout the body including the skin lung liver and in the context of cancer cells. MMP activity is important for normal physiological processes like tissue remodeling and wound healing. Researchers frequently use MMP activity assay kits to measure the activity levels of these enzymes in biological samples for both research and clinical diagnostics.
Biological function summary
MMPs play significant roles in tissue remodeling and cell signaling. MMPs are not usually found as part of a complex but individual enzymes can be regulated through inhibitor proteins such as TIMPs (tissue inhibitors of metalloproteinases) which bind to MMPs and modulate their enzymatic activity. This regulation is essential to maintain tissue homeostasis and any imbalance may result in pathological conditions. MMPs influence processes such as morphogenesis angiogenesis and metastasis highlighting their importance in developmental biology and cancer research.
Pathways
MMPs are integral to processes involving the breakdown of the extracellular matrix. Notably MMPs are central components of the degradative pathways facilitating cell migration during immune responses and metastasis. MMPs interact closely with protein inhibitors like TIMPs and these interactions play key roles in the balance of proteolytic activity. Additionally MMPs relate to other proteins like integrins and cytokines linking them to pathways such as the TGF-beta signaling pathway and the Wnt signaling pathway.
MMPs have been implicated in conditions such as cancer and arthritis. In cancer abnormal MMP activity can elevate tumor invasion and metastasis where MMPs collaborate with proteins like E-cadherin and VEGF modulating the tumor microenvironment. In arthritis MMPs contribute to the degradation of cartilage intensifying inflammatory responses and tissue damage. Understanding MMP function and its inhibition offers therapeutic potential for managing these diseases.
