Cathepsin L Activity Fluorometric Assay Kit

• Cat.No.: Kit-0186
• Product Overview: Cathepsin L Activity Assay Kit is a homogeneous assay that can be used to detect the activity of enzyme in biological samples or in purified enzyme preparations. The long wavelength FRET substrate is designed based on a sequence surrounding the cleavage site of Cathepsin L. A Fluor 488 pair is used for optimal quenching of the intact substrate. When active Cathepsin L cleaves the FRET substrate, it results in an increase of Fluor 488 fluorescence and can be monitored at excitation/emission = 490 nm/520 nm. The fluorescent signal from Fluor 488 is stable at low pH, the optimal pH for Cathepsin L activity.

Specification

• Tag: Non
• Size: 100 Assays
• Description: Apoptosis can be mediated by mechanisms other than the traditional caspase-mediated cleavage cascade. There is growing recognition that alternative proteolytic enzymes such as the lysosomal cathepsin proteases may initiate or propagate proapoptotic signals. Cathepsins are lysosomal enzymes that are also used as sensitive markers in various toxicological investigations. The Cathepsin-L Activity Assay kit is a fluorescence-based assay that utilizes the preferred cathepsin-L substrate sequence FR labeled with AFC (amino-4-trifluoromethyl coumarin). Cell lysates or other samples that contain cathepsin-L will cleave the synthetic substrate FR-AFC to release free AFC.
• Applications: Detects alternative proteolytic enzymes such as the lysosomal cathepsin proteases that can initiate or propagate proapoptotic signals.
• Target Species: Mammalian
• Usage: For Research Use Only! Not For Use in Humans.
• Storage: Store component C at -80°C. Store all other components at -20°C. Component D can be stored at room temperature for convenience. Protect components A and B from light and moisture.
• Kit Components: Component A: Cathepsin L substrate, Ex/Em=490 nm/520 nm upon cleavage: 1 mM, 50 µL Component B: Fluorescence reference standard, Ex/Em=490 nm/520 nm: 1 mM, 10 µL Component C: Cathepsin L, human recombinant: 0.1 mg/mL, 20 µL Component D: Assay Buffer: 20 mL Component E: Cathepsin L inhibitor: 100 µM, 10 µL Component F: DTT: 1 M, 200 µL
• Detection method: Fluorescence (Ex/Em 400/505)
• Compatible Sample Types: Cell and tissue lysates
• Shipping: Dry ice
• Materials Required but Not Supplied: 96-well microplate: Black, flat bottom 96-well plate with non-binding surface. Fluorescence microplate reader: Capable of detecting emission at 520 nm with excitation at 490 nm.