Recombinant Human FAP His-tag (Mammalian) Protein, CF

R&D Systems | Catalog # 10921-SE

Analyzed by SEC-MALS.
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Key Product Details

  • R&D Systems CHO-derived Recombinant Human FAP His-tag (Mammalian) Protein (10921-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

Q12884.5

Applications

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

Product Specifications

Source

Chinese Hamster Ovary cell line, CHO-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

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

N-terminal Sequence Analysis

His

Predicted Molecular Mass

86 kDa

SDS-PAGE

83-92 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 Human FAP His-tag (Mammalian) Protein, CF

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

Recombinant Human FAP His-tag (Mammalian) (Catalog # 10921-SE) has a molecular weight (MW) of 194.0 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 His-tag (Mammalian) Protein Enzyme Activity

Recombinant Human FAP His-tag (Mammalian) (Catalog # 10921-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).

Recombinant Human FAP His-tag Protein SDS-PAGE.

2 μg/lane of Recombinant Human FAP His-tag Protein (Catalog # 10921-SE) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 83-92 kDa.

Formulation, Preparation, and Storage

10921-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). 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

Q12884.5

Additional Fibroblast Activation Protein alpha/FAP Products

Product Documents for Recombinant Human FAP His-tag (Mammalian) Protein, CF

Certificate of Analysis

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Product Specific Notices for Recombinant Human FAP His-tag (Mammalian) Protein, CF

For research use only

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Protocols

View specific protocols for Recombinant Human FAP His-tag (Mammalian) Protein, CF (10921-SE):

Materials
  • Assay Buffer: 50 mM Tris, 1 M NaCl, 1 mg/mL BSA, pH 7.5
  • Recombinant Human FAP (rhFAP) (Catalog # 10921-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 M5 by Molecular Devices) or equivalent
  1. Dilute rhFAP to 0.2 µg/mL in Assay Buffer. 
  2. Dilute Substrate to 200 µM in Assay Buffer.
  3. Load in a plate 50 µL of 0.2 µg/mL of rhFAP, and start the reaction by adding 50 µL of 200 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 Substrate Blank
**Derived using calibration standard 7-Amino, 4-Methyl Coumarin (Sigma, Catalog # A-9891).

Per Well:

  • rhFAP: 0.01 µg
  • Substrate: 100 µM



























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