Recombinant Human FAP Protein, CF

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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).

Product Details

Summary
Reactivity HuSpecies Glossary
Applications Enzyme Activity
Format
Carrier-Free

Order Details

Recombinant Human FAP Protein, CF Summary

Details of Functionality
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.
Source
Spodoptera frugiperda, Sf 21 (baculovirus)-derived human Fibroblast Activation Protein alpha/FAP protein
Leu26-Asp760, with an N-terminal 6-His tag
Accession #
N-terminal Sequence
His
Protein/Peptide Type
Recombinant Enzymes
Gene
FAP
Purity
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Note
<1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

Dilutions
  • Enzyme Activity
Theoretical MW
86 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.
SDS-PAGE
85 kDa, reducing conditions
Publications
Read Publications using
3715-SE in the following applications:

Packaging, Storage & Formulations

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.
Buffer
Supplied as a 0.2 μm filtered solution in Tris, NaCl and Glycerol.
Purity
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Assay Procedure
  • 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

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human FAP Protein, CF

  • 170 kDa melanoma membrane-bound gelatinase
  • DKFZp686G13158
  • DPPIV
  • EC 3.4.21.-
  • FAP
  • FAPA
  • Fibroblast Activation Protein alpha
  • fibroblast activation protein, alpha
  • Integral membrane serine protease
  • Seprase
  • vibronectin

Background

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).
  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.

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Publications for Fibroblast Activation Protein alpha/FAP (3715-SE)(14)

We have publications tested in 2 confirmed species: Human, N/A.

We have publications tested in 7 applications: Binding Assay, Bioassay, ELISA (Standard), Enzyme Assay, Reference Standard, Standard, Western Blot.


Filter By Application
Binding Assay
(1)
Bioassay
(6)
ELISA (Standard)
(1)
Enzyme Assay
(2)
Reference Standard
(1)
Standard
(1)
Western Blot
(1)
All Applications
Filter By Species
Human
(11)
N/A
(3)
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Showing Publications 1 - 10 of 14. Show All 14 Publications.
Publications using 3715-SE Applications Species
H Ren, XZ Zeng, XX Zhao, DY Hou, H Yao, M Yaseen, L Zhao, WH Xu, H Wang, LL Li A bioactivated in vivo assembly nanotechnology fabricated NIR probe for small pancreatic tumor intraoperative imaging Nature Communications, 2022;13(1):418. 2022 [PMID: 35058435] (Bioassay, Human) Bioassay Human
CR Xi, A Di Fazio, NA Nadvi, MSW Xiang, HE Zhang, C Deshpande, Y Chen, MS Tabar, XM Wang, CG Bailey, GW McCaughan, WB Church, MD Gorrell An improved production and purification protocol for recombinant soluble human fibroblast activation protein alpha Protein Expression and Purification, 2021;181(0):105833. 2021 [PMID: 33524496] (Human) Human
JM Cho, EH Yang, W Quan, EH Nam, HG Cheon Discovery of a novel fibroblast activation protein (FAP) inhibitor, BR103354, with anti-diabetic and anti-steatotic effects Scientific Reports, 2020;10(1):21280. 2020 [PMID: 33277568] (Bioassay, N/A) Bioassay N/A
H Wei, Y Xu, Y Wang, L Xu, C Mo, L Li, B Shen, Y Sun, P Cheng, L Yang, Y Pang, A Qin, Y Cao, SJ Morrison, R Yue Identification of Fibroblast Activation Protein as an Osteogenic Suppressor and Anti-osteoporosis Drug Target Cell Rep, 2020;33(2):108252. 2020 [PMID: 33053358] (Bioassay, Human) Bioassay Human
X Wang, F Gessier, R Perozzo, D Stojkov, A Hosseini, K Amirshahro, S Kuchen, S Yousefi, P Lötscher, HU Simon RIPK3-MLKL-Mediated Neutrophil Death Requires Concurrent Activation of Fibroblast Activation Protein-alpha J. Immunol., 2020;0(0):. 2020 [PMID: 32796025] (Reference Standard, Human) Reference Standard Human
R Wang, C Zhang, C Zheng, H Li, X Xie, Y Jin, Z Liu, H Chen Introduction of Z-GP scaffold into procarbazine reduces spermatoxicity and myelosuppression Bioorg. Chem., 2018;83(0):461-467. 2018 [PMID: 30448724] (Enzyme Assay, Human) Enzyme Assay Human
XP Chai, GL Sun, YF Fang, LH Hu, X Liu, XW Zhang Tumor-targeting efficacy of a BF211 prodrug through hydrolysis by fibroblast activation protein-? Acta Pharmacol. Sin., 2017;0(0):. 2017 [PMID: 29119969] (Bioassay, Human) Bioassay Human
M Chen, X Lei, C Shi, M Huang, X Li, B Wu, Z Li, W Han, B Du, J Hu, Q Nie, W Mai, N Ma, N Xu, X Zhang, C Fan, A Hong, M Xia, L Luo, A Ma, H Li, Q Yu, H Chen, D Zhang, W Ye Pericyte-targeting prodrug overcomes tumor resistance to vascular disrupting agents J. Clin. Invest., 2017;0(0):. 2017 [PMID: 28846068] (Enzyme Assay, Human) Enzyme Assay Human
S Uitte de W, FM Keane, DG Bowen, JJMC Malfliet, HE Zhang, B Maneck, GW McCaughan, FWG Leebeek, DC Rijken, MD Gorrell Circulating fibroblast activation protein activity and antigen levels correlate strongly when measured in liver disease and coronary heart disease PLoS ONE, 2017;12(6):e0178987. 2017 [PMID: 28582421] (ELISA (Standard), Human) ELISA (Standard) Human
The Expression of Fibroblast Activation Protein in Clear Cell Renal Cell Carcinomas Is Associated with Synchronous Lymph Node Metastases PLoS ONE, 2016;11(12):e0169105. 2016 [PMID: 28033421] (Western Blot, Human) Western Blot Human
Show All 14 Publications.

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Bioinformatics

Gene Symbol FAP
Uniprot