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Recombinant Human FAP Protein, CF

R&D Systems | Catalog # 3715-SE

New Version Available: 10921-SE
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Analyzed by SEC-MALS
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Key Product Details

  • R&D Systems Sf 21 (baculovirus)-derived Recombinant Human FAP Protein (3715-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

Sf 21 (baculovirus)

Accession Number

Q12884

Applications

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

Product Specifications

Source

Spodoptera frugiperda, Sf 21 (baculovirus)-derived human Fibroblast Activation Protein alpha/FAP protein
Leu26-Asp760, with an N-terminal 6-His tag

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

His

Predicted Molecular Mass

86 kDa

SDS-PAGE

85 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 >1800 pmol/min/µg, as measured under the described conditions.

Scientific Data Images for Recombinant Human FAP Protein, CF

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

Recombinant Human FAP/His (Catalog # 3715-SE) has a molecular weight (MW) of 176.3 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 Human FAP Protein Enzyme Activity

Recombinant Human FAP Protein Enzyme Activity

Recombinant Human FAP His-tag (Catalog # 3715-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

3715-SE
Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl and Glycerol.
Shipping The product is shipped with dry ice or equivalent. 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, -70 °C as supplied.
  • 3 months, -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 (ECD), human FAP shares 90% amino acid (aa) sequence identity with mouse and rat FAP (3, 4). Alternative splicing of human FAP generates a truncated isoform that consists of the C-terminal 239 residues of the ECD. A soluble and enzymatically active form of FAP known as antiplasmin-cleaving enzyme (APCE) circulates in human plasma (4). FAP is expressed on reactive stromal fibroblasts in tumor tissue and wound healing and on synoviocytes in rheumatoid arthritis (1, 6-8). It exhibits dipeptidyl peptidase activity with substrate specificity similar to DPPIV, which is specific for N-terminal Xaa-Pro sequences (6, 9). FAP is also an endopeptidase that can degrade Gelatin, Collagens I and IV, Fibronectin, and Laminin (1, 6, 9) as well as several peptide hormones (e.g. Neuropeptide Y, Brain Natriuretic Peptide, Substance P, Peptide YY, and Incretins) (10). The enzymatic activity is dependent on FAP association with DPPIV on the cell surface (6, 9, 11, 12). The matrix-dedgrading activity of FAP contributes to tumor cell migration and invasion (11-14). In addition, FAP can enhance tumor cell growth by limiting the development of anti-tumor immunity (15).

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

Q12884

Additional Fibroblast Activation Protein alpha/FAP Products

Product Documents for Recombinant Human FAP Protein, CF

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Recombinant Human FAP Protein, CF

For research use only

Citations for Recombinant Human FAP Protein, CF

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Protocols

View specific protocols for Recombinant Human FAP Protein, CF (3715-SE):

Materials
  • Assay Buffer: 50 mM Tris, 1 M NaCl, 1 mg/mL BSA, pH 7.5
  • Recombinant Human Fibroblast Activation Protein alpha /FAP (rhFAP) (Catalog # 3715-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 rhFAP 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 rhFAP 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 # A-9891).

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

FAQs for Recombinant Human FAP Protein, CF

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    • A: Recombinant Human FAP Protein, CF (Catalog # 3715-SE) contains an N-terminal 6-His tag. Please refer to the Source section on the product-specific page for sequence information.
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