FAP Fluorogenic Assay Kit

Catalog #
80210
$1,190 *
Size: 96 reactions
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*US Pricing only. For international pricing, please contact your local distributor.
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Description

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. 

Need us to run inhibitor screens or profile your compounds against FAP? Check out our Protease Screening Services.

This product has been cited 6 times.

Synonyms
Prolyl endopeptidase FAP, 170 kDa melanoma membrane-bound gelatinase, Dipeptidyl peptidase FAP, Fibroblast activation protein alpha, FAPalpha, Gelatine degradation protease FAP, Integral membrane serine protease, Post-proline cleaving enzyme
Product Info
Storage and Usage
Citations6
Assay Kit Format
Fluorogenic
Materials Required But Not Supplied

Fluorescent microtiter plate reader capable of measurement at λex350-380/λem440-460 nm

Format
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.

UniProt #
Q12884
Background

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.

References

Pure E. and Blomberg R., 2018, Oncogene 37: 4343-4357.
Bughda R., et al., 2021, Immunotargets Ther. 10:313-323.