FAP Fluorogenic Assay Kit
The FAP Fluorogenic Assay Kit is designed to measure FAP (fibroblast activation protein) protease activity for screening and profiling applications. It comes in a convenient 96-well format, with purified recombinant FAP enzyme (amino acids 29-760), DPP substrate, and DPP assay buffer for 100 enzyme reactions.
The DPP fluorogenic substrate is incubated with a sample containing FAP enzyme to produce a fluorophore that can then be measured using a fluorescence reader.
Figure 1: Illustration of the assay principle.
Fluorogenic DPP Substrate 1 is a fluorogenic peptide substrate (Ala-Pro-AMC dipeptide). In the conjugated form the energy emitted from the fluorochrome AMC is quenched. Proteolysis releases AMC and fluorescence is emitted. The increase in fluorescence is directly proportional to FAP activity.
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This product has been cited 6 times.
Fluorescent microtiter plate reader capable of measurement at λex350-380/λem440-460 nm
Catalog # | Name | Amount | Storage |
80100 | FAP, His-Tag* | 30 µg | -80°C |
80300 | DPP Assay Buffer | 10 ml | -20°C |
80305 | 0.5 mM Fluorogenic DPP Substrate 1 | 100 μl | -80°C |
50 µM AMC Fluorescent Standard | 500 μl | -20°C | |
79685 | 96-well black microplate | 1 | Room Temp |
* The concentration of protein is lot-specific and will be indicated on the tube containing the protein.
Fibroblast Activation Protein, or FAP, is a type II membrane serine protease of the dipeptidyl peptidase (DPP) subfamily. It is involved in wound healing and tissue repair. FAP upregulation is seen on activated stromal fibroblasts present in cancer, including breast, lung, prostate, pancreatic and cervical cancer. It has an impact on the tumor microenvironment (TME), where it reduces the levels of PEDF (pigment epithelium derived factor), angiopoietin-1 and VEGF-c (vascular endothelial growth factor c) and increase TGF-β (transforming growth factor β) and create an immunosuppressive environment. It is used as biomarker for pro-tumorigenic stroma and links to a poor prognosis. Studies in animal models have shown that knock-out of FAP reduces the metastasis of pancreatic ductal adenocarcinomas (PDA). It has thus been proposed as a candidate target for small molecule inhibitors for cancer therapy applications. Additionally, its expression profile makes it a good candidate for CAR-T applications, with pre-clinical and clinical trials currently ongoing.
Pure E. and Blomberg R., 2018, Oncogene 37: 4343-4357.
Bughda R., et al., 2021, Immunotargets Ther. 10:313-323.