Recombinant Cynomolgus Fibroblast Activation Protein alpha

R&D Systems | Catalog # 10278-SE

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Key Product Details

  • R&D Systems CHO-derived Recombinant Cynomolgus Fibroblast Activation Protein alpha (10278-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

CHO

Accession Number

XP_005573377

Applications

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

Product Specifications

Source

Chinese Hamster Ovary cell line, CHO-derived cynomolgus monkey Fibroblast Activation Protein alpha/FAP protein
Leu26-Asp760

Purity

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

Endotoxin Level

<0.10 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Leu26

Predicted Molecular Mass

85 kDa

SDS-PAGE

87-94 kDa, under 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 >2500 pmol/min/μg, as measured under the described conditions.

Scientific Data Images for Recombinant Cynomolgus Fibroblast Activation Protein alpha

Recombinant Cynomolgus Fibroblast Activation Protein alpha Enzyme Activity

Recombinant Cynomolgus Fibroblast Activation Protein alpha Enzyme Activity

Recombinant Cynomolgus Fibroblast Activation Protein (Catalog # 10278-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-methlycoumarin (AMC).

Formulation, Preparation, and Storage

10278-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 forms a homodimer and is structurally related to the dipeptidyl peptidase IV (DPPIV/CD26) family with a short cytoplasmic tail, a single transmembrane domain, and an extracellular domain that contains the active site (1-3). Within the extracellular domain, cynomolgous FAP shares 99.6% and 89.8% amino acid (aa) sequence identity with human and mouse FAP, respectively. It exhibits dipeptidyl peptidase activity with substrate specificity similar to DPPIV, which is specific for N-terminal Xaa-Pro sequences (4, 5). FAP is also an endopeptidase that can degrade Gelatin, Collagens I and IV, Fibronectin, and Laminin (1, 4, 5) as well as several peptide hormones (e.g. Neuropeptide Y, Brain Natriuretic Peptide, Substance P, Peptide YY, and Incretins) (6). FAP is also known to interact with several surface molecules to play roles in cell signaling, cell invasion and migration (3). Although not detectible in normal tissues, FAP is elevated in activated stromal fibroblasts, tumor-associated macrophages, activated hepatic stellate cells and myofibroblasts during pathological conditions that include tissue remodeling such as most types of cancer, wound healing, arthritis, atherosclerosis, and fibrosis (1, 3, 4, 7-9). Targeting FAP with vaccines, antibodies, or pharmacologics impairs tumor progression in several cancer models making it a promising immunotherapy target (9-12).

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. Lay, A.J. et. al. (2019) Front. Biosci. 24:1.
  4. Park, J.E. et al. (1999) J. Biol. Chem. 274:36505.
  5. Aertgeerts, K. et al. (2005) J. Biol. Chem. 280:19441.
  6. Keane, F.M. et al. (2011) FEBS J. 278:1316.
  7. Rettig, W.J. et al. (1988) Proc. Natl. Acad. Sci. USA 85:3110.
  8. Bauer, S. et al. (2006) Arthritis Res. 8:R171.
  9. Jiang, G.M. et al. (2016) Oncotarget 7:33472.
  10. Cheng, J.D. et al. (2002) Cancer Res. 62:4767.
  11. Kraman, M. et al. (2010) Science 330:827.
  12. Busek, P. et al. (2018) Front. Biosci. 23:1933.

Alternate Names

FAP, Seprase

Entrez Gene IDs

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

Gene Symbol

FAP

UniProt

XP_005573377

Additional Fibroblast Activation Protein alpha/FAP Products

Product Documents for Recombinant Cynomolgus Fibroblast Activation Protein alpha

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 Cynomolgus Fibroblast Activation Protein alpha

For research use only

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Protocols

View specific protocols for Recombinant Cynomolgus Fibroblast Activation Protein alpha (10278-SE):

Materials
  • Assay Buffer: 50 mM Tris, 1 M NaCl, 1 mg/mL BSA, pH 7.5
  • Recombinant Cynomolgus Monkey Fibroblast Activation Protein  alpha /FAP (cynoFAP) (Catalog # 10278-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 rcynoFAP to 0.2 µg/mL in Assay Buffer.
  2. Dilute Substrate to 100 µM in Assay Buffer.
  3. Load in plate 50 µL of 0.2 µg/mL rcynoFAP, and start the reaction by adding 50 µL of 100 µM Substrate. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of 100 µM Substrate.
  4. Read at excitation and emission wavelengths of 380 nm and 460 nm (top read), respectively, in kinetic mode of 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 7-amino, 4-Methyl Coumarin (Sigma, Catalog # A9891).

Per Well:
  • rcynoFAP: 0.01 µg
  • Substrate: 50 µM

FAQs for Recombinant Cynomolgus Fibroblast Activation Protein alpha

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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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