SensoLyte® 520 Cathepsin E Assay Kit Fluorimetric, AnaSpec, Inc.
Supplier: Anaspec Inc
Cathepsin E is a non-lysosomal aspartic proteinase of the pepsin superfamily, playing an important role in host defense against cancer cells in addition to it’s roles in protein degradation, antigen processing etc
Cathepsins are a class of globular lysosomal proteases playing a vital role in mammalian cellular turnover. They degrade polypeptides and are distinguished by their substrate specificities.
The cathepsin assay kits offers a series of fluorimetric long wavelength, sensitive SensoLyte® 520 assay kits for enzyme activity determination of Cathepsins G, D, E, B, K, L and S. These kits employ a FRET (Fluorescence Resonance Energy Transfer) peptide substrate that emits in the green range of 520 nm, with excitation at 490 nm. The advantage of using longer wavelength emitting substrates means that autofluorescence from cellular components and test compounds is minimal; thus making the kits more sensitive. Furthermore, the use of HiLyte Fluor™ 488 (except for Cathepsin S, which uses 5-FAM) as the FRET donor (fluorophore) results in a more stable fluorescent signal, since HiLyte Fluor™ 488 is resistant to acidic pH. A listing of the SensoLyte® Cathepsin Kits providing conventional FRET (Mca/Dnp) or fluorogenic substrates for cathepsins are also available.
Kits detect specific Cathepsin activities; provides ample reagents to perform 100 assays in 96-well format; minimal hands-on time required; detailed protocol and references provided.
SensoLyte® 520 Cathepsin E Activity Assay Kit is a homogeneous assay used to detect the enzyme activity in biological samples or in purified enzyme preparations and features a unique long wavelength FRET substrate to reduce the cross reactivity of Cathepsin D. A QXL® 520/HiLyte™ Fluor 488 pair is used for optimal quenching of the intact substrate. When active Cathepsin E cleaves the FRET substrate, it results in an increase of HiLyte™ Fluor 488 fluorescence, monitored at excitation/emission = 490 nm/520 nm. The fluorescent signal from HiLyte™ Fluor 488 is stable at low pH, an optimal pH for cathepsin activity. This assay can detect as low as 2 ng/mL active Cathepsin E.
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