Anaspec has introduced the Sensolyte 520 MMP-15, 16 and 24 assay kits for high-throughput screening of inducers and inhibitors for MMP-15, MMP-16 and MMP-24.
The substrate used in each of these kits detects MMP-15, 16 or 24 activity using a 5-FAM/QXL 520 FRET peptide.
In the intact FRET peptide, the fluorescence of 5-FAM is quenched by QXL 520.
Upon cleavage into two separate fragments by MMP-15, 16 or 24, the fluorescence of 5-FAM is recovered, and can be monitored at excitation/emission wavelengths = 490nm/520nm.
With good fluorescence quantum yield and longer wavelength, 5-FAM shows less interference from auto-fluorescence of test compounds and cellular components.
The assays are performed in a 96-well microplate format.
The 5-FAM/QXL 520 substrates can detect sub-nanogram levels of MMP-15, 16 and 24 activity.
Anaspec has also introduced the Sensolyte 390 generic MMP activity assay kit, which is optimised to detect the activity of a variety of MMPs, including MMP-1, 2, 7, 8, 9, 13, 14, 15, 16, and 24.
It can be used for detecting generic MMP activity in biological samples or for high-throughput screening of MMP inducers and inhibitors using purified MMPs.
An Mca/Dnp FRET peptide is used as a substrate.
In the intact FRET peptide, the fluorescence of Mca is quenched by Dnp.
Upon cleavage into two separate fragments by MMPs, the fluorescence of Mca is recovered, and can be monitored at Ex/Em = 330/390nm.
The assays are also performed in a 96-well microplate format.
The matrix metalloproteinases (MMPs) constitute a family of zinc-dependent endopeptidases that function within the extracellular matrix (ECM).
These enzymes are responsible for the breakdown of connective tissues and are important in bone remodelling, the menstrual cycle, and repair of tissue damage.
MMP-15 (also known as MT2-MMP), MP-16 (MT3-MMP) and MMP-24 (MT5-MMP) are members of the membrane-type MMP (MT-MMP) subfamily.
MMP-15 contains a potential transmembrane domain and is expressed at the cell surface rather than secreted.
MMP-16 is found in soluble form, while MMP-24 may function as a soluble proteinase since it is shed from cell membrane.
All these three MMPs activate pro-MMP-2 (gelatinase-A) that is involved in tumour development and metastasis, and thus are proposed as therapeutic targets for cancer.
