Recombinant Mouse FAP His-tag (Catalog # 8647-SE) is measured by its ability to convert the substrate benzyloxycarbonyl-Gly-Pro-7-amido-4-methylcoumarin (Z-GP-AMC) to Z-Gly-Pro and 7-amino-4-methylcoumarin (AMC).
Measured by its ability to convert the substrate benzyloxycarbonyl-Gly-Pro-7-amido-4-methylcoumarin (Z-GP-AMC) to Z-Gly-Pro and 7-amino-4-methylcoumarin (AMC). The specific activity is >2000 pmol/min/μg, as measured under the described conditions.
Mouse myeloma cell line, NS0-derived mouse Fibroblast Activation Protein alpha/FAP protein Leu26-Asp761
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
<1.0 EU per 1 μg of the protein by the LAL method.
85 kDa. Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Substrate: Z-Gly-Pro-AMC (Bachem, Catalog # I-1145), 10 mM stock in DMSO
F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
Dilute rmFAP to 0.2 µg/mL in Assay Buffer.
Dilute Substrate to 100 µM in Assay Buffer.
Load 50 µL of 0.2 µg/mL of rmFAP into a plate, and start the reaction by adding 50 µL of 100 µM Substrate. Include a Substrate Blank containing 50 µL of Assay Buffer and 50 µL of Substrate.
Read at excitation and emission wavelengths of 380 nm and 460 nm (top read), respectively, in kinetic mode for 5 minutes.
Calculate specific activity:
Specific Activity (pmol/min/µg) =
Adjusted Vmax* (RFU/min) x Conversion Factor** (pmol/RFU)
amount of enzyme (µg)
*Adjusted for Substrate Blank **Derived using calibration standard 7-Amino, 4-Methyl Coumarin (Sigma, Catalog # A9891).
rmFAP: 0.010 µg
Substrate: 50 µM
This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.
Alternate Names for Recombinant Mouse FAP Protein, CF
170 kDa melanoma membrane-bound gelatinase
Fibroblast Activation Protein alpha
fibroblast activation protein, alpha
Integral membrane serine protease
FAP (also known as seprase) is a 95 kDa Type II transmembrane serine protease that is structurally related to dipeptidyl peptidase IV (DPPIV/CD26) (1, 2). Within the extracellular domain, mouse FAP shares 90% and 97% amino acid (aa) sequence identity with human and rat FAP, respectively (3, 4). Alternative splicing of mouse FAP generates isoforms with a 33 aa or 5 aa deletion in the extracellular juxtamembrane region (3). FAP is expressed on reactive stromal fibroblasts in tumor tissue and wound healing and on synoviocytes in rheumatoid arthritis (1, 5-7). It exhibits dipeptidyl peptidase activity with substrate specificity similar to DPPIV, which is specific for N-terminal Xaa-Pro sequences (5, 8). FAP is also an endopeptidase that can degrade Gelatin, Collagens I and IV, Fibronectin, and Laminin (1, 5, 8) as well as several peptide hormones (e.g. Neuropeptide Y, Brain Natriuretic Peptide, Substance P, Peptide YY, and Incretins) (9). The enzymatic activity is dependent on FAP association with DPPIV on the cell surface (5, 8, 10, 11). The matrix-dedgrading activity of FAP contributes to tumor cell migration and invasion (10-13). In addition, FAP can enhance tumor cell growth by limiting the development of anti-tumor immunity (14).
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Bauer, S. et al. (2006) Arthritis Res. 8:R171.
Aertgeerts, K. et al. (2005) J. Biol. Chem. 280:19441.
Keane, F.M. et al. (2011) FEBS J. 278:1316.
Ghersi, G. et al. (2006) Cancer Res. 66:4652.
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