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Abcam's 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, which can be measured in a spectrophotometer at Em = 505 nm. The amount of free AFC will be directly proportional to the cathepsin L activity found in the sample.
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Cathepsin L plays a major role in antigen processing, tumor invasion and metastasis, bone resorption, and turnover of intracellular and secreted proteins involved in growth regulation. Although commonly recognized as a lysosomal protease, cathepsin L is also secreted. This broad-spectrum protease is potent in degrading several extracellular proteins (laminins, fibronectin, collagens I and IV, elastin, and other structural proteins of basement membranes) as well as serum proteins and cytoplasmic and nuclear proteins.
If you need additional Ac-FR-AFC substrate (10mM), it is avaible for purchasing as ab157769.
|Assay Buffer||1 x 30ml|
|CL Inhibitor (1 mM)||Red||1 x 20µl|
|CL Positive Control (human)||1 vial|
|CL Substrate Ac-FR-AFC (10 mM)||Brown||1 x 200µl|
|DTT||1 x 100µl|
Our Abpromise guarantee covers the use of ab65306 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Functional Studies||Use at an assay dependent dilution.|
Cathepsin L activity in cell lysates (duplicates +/- SD). Data from lysates corresponding to 0.5-2 million cells per well.
Cathepsin L activity in biological fluids (serum in the range of neat-1/27 dilution, plasma neat; duplicates +/- SD)
Image obtained from Stratigopoulos G et al; JBC, 2011 Jan 21;286(3):2155-70