Recombinant Mouse FAP Protein, CF

R&D Systems | Catalog # 8647-SE

Analyzed by SEC-MALS
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Key Product Details

  • R&D Systems NS0-derived Recombinant Mouse FAP Protein (8647-SE)
  • Quality control testing to verify active proteins with lot specific assays by in-house scientists
  • All R&D Systems proteins are covered with a 100% guarantee

Source

NS0

Accession Number

P97321

Applications

Enzyme Activity
View all Fibroblast Activation Protein alpha/FAP Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived mouse Fibroblast Activation Protein alpha/FAP protein
Leu26-Asp761

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<1.0 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Leu26 & Val31

Predicted Molecular Mass

85 kDa

SDS-PAGE

78-90 kDa, reducing conditions

Activity

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.

Scientific Data Images for Recombinant Mouse FAP Protein, CF

Recombinant Mouse Fibroblast Activation Protein alpha /FAP Protein SEC-MALS.

Recombinant Mouse FAP (Catalog # 8647-SE) has a molecular weight (MW) of 173.8 kDa as analyzed by SEC-MALS, suggesting that this protein is a homodimer.  MW may differ from predicted MW due to post-translational modifications (PTMs) present (i.e. Glycosylation).
Recombinant Mouse FAP Protein Enzyme Activity

Recombinant Mouse FAP Protein Enzyme Activity

Recombinant Mouse FAP (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).

Formulation, Preparation, and Storage

8647-SE
Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl and Glycerol.
Shipping The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.

Background: Fibroblast Activation Protein alpha/FAP

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).

References

  1. Zi, F. et al. (2015) Mol. Med. Rep. 11:3203.
  2. Pineiro-Sanchez, M.L. et al. (1997) J. Biol. Chem. 272:7595.
  3. Niedermeyer, J. et al. (1997) Int. J. Cancer 71:383.
  4. Scanlan, M.J. et al. (1994) Proc. Natl. Acad. Sci. USA 91:5657.
  5. Park, J.E. et al. (1999) J. Biol. Chem. 274:36505.
  6. Rettig, W.J. et al. (1988) Proc. Natl. Acad. Sci. USA 85:3110.
  7. Bauer, S. et al. (2006) Arthritis Res. 8:R171.
  8. Aertgeerts, K. et al. (2005) J. Biol. Chem. 280:19441.
  9. Keane, F.M. et al. (2011) FEBS J. 278:1316.
  10. Ghersi, G. et al. (2006) Cancer Res. 66:4652.
  11. Ghersi, G. et al. (2002) J. Biol. Chem. 277:29231.
  12. Cheng, J.D. et al. (2005) Mol. Cancer Ther. 4:351.
  13. Cheng, J.D. et al. (2002) Cancer Res. 62:4767.
  14. Kraman, M. et al. (2010) Science 330:827.

Alternate Names

FAP, Seprase

Entrez Gene IDs

2191 (Human); 14089 (Mouse); 102134935 (Cynomolgus Monkey)

Gene Symbol

FAP

UniProt

P97321

Additional Fibroblast Activation Protein alpha/FAP Products

Product Documents for Recombinant Mouse FAP Protein, CF

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Recombinant Mouse FAP Protein, CF

For research use only

Citations for Recombinant Mouse FAP Protein, CF

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Protocols

View specific protocols for Recombinant Mouse FAP Protein, CF (8647-SE):

Materials
  • Assay Buffer: 50 mM Tris, 1 M NaCl, 1 mg/mL BSA, pH 7.5
  • Recombinant Mouse Fibroblast Activation Protein alpha /FAP (rmFAP) (Catalog # 8647-SE)
  • 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
  1. Dilute rmFAP to 0.2 µg/mL in Assay Buffer.
  2. Dilute Substrate to 100 µM in Assay Buffer.
  3. 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.
  4. Read at excitation and emission wavelengths of 380 nm and 460 nm (top read), respectively, in kinetic mode for 5 minutes.
  5. 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).

Per Well:
  • rmFAP: 0.010 µg
  • Substrate: 50 µM

FAQs for Recombinant Mouse FAP Protein, CF

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  • Q: Does this enzyme have a tag?

    A: No, this enzyme does not have a tag.  Please refer to the Source section on the product-specific page or product datasheet for sequence information.

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