Recombinant Human FAP Protein, CF

Catalog # Availability Size / Price Qty
3715-SE-010
Recombinant Human FAP Protein Enzyme Activity
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Product Details
Citations (14)
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Recombinant Human FAP Protein, CF Summary

Product Specifications

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.
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.
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
Analysis
His
Predicted Molecular Mass
86 kDa
SDS-PAGE
85 kDa, reducing conditions

Product Datasheets

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3715-SE

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

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.

Assay Procedure

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

Scientific Data

Enzyme Activity Recombinant Human FAP Protein Enzyme Activity View Larger

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).

Reconstitution Calculator

Reconstitution Calculator

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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.
Entrez Gene IDs
2191 (Human); 14089 (Mouse); 102134935 (Cynomolgus Monkey)
Alternate Names
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

Citations for Recombinant Human FAP Protein, CF

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

14 Citations: Showing 1 - 10
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  1. A bioactivated in vivo assembly nanotechnology fabricated NIR probe for small pancreatic tumor intraoperative imaging
    Authors: H Ren, XZ Zeng, XX Zhao, DY Hou, H Yao, M Yaseen, L Zhao, WH Xu, H Wang, LL Li
    Nature Communications, 2022;13(1):418.
    Species: Human
    Sample Types: Recombinant Proteins
    Applications: Bioassay
  2. An improved production and purification protocol for recombinant soluble human fibroblast activation protein alpha
    Authors: 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
    Protein Expression and Purification, 2021;181(0):105833.
    Species: Human
    Sample Types: Reference Standard
  3. Discovery of a novel fibroblast activation protein (FAP) inhibitor, BR103354, with anti-diabetic and anti-steatotic effects
    Authors: JM Cho, EH Yang, W Quan, EH Nam, HG Cheon
    Scientific Reports, 2020;10(1):21280.
    Species: N/A
    Sample Types:
    Applications: Bioassay
  4. Identification of Fibroblast Activation Protein as an Osteogenic Suppressor and Anti-osteoporosis Drug Target
    Authors: 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
    Cell Rep, 2020;33(2):108252.
    Species: Human
    Sample Types: Reference Standard
    Applications: Bioassay
  5. RIPK3-MLKL-Mediated Neutrophil Death Requires Concurrent Activation of Fibroblast Activation Protein-alpha
    Authors: X Wang, F Gessier, R Perozzo, D Stojkov, A Hosseini, K Amirshahro, S Kuchen, S Yousefi, P Lötscher, HU Simon
    J. Immunol., 2020;0(0):.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Reference Standard
  6. Introduction of Z-GP scaffold into procarbazine reduces spermatoxicity and myelosuppression
    Authors: R Wang, C Zhang, C Zheng, H Li, X Xie, Y Jin, Z Liu, H Chen
    Bioorg. Chem., 2018;83(0):461-467.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Enzyme Assay
  7. Pericyte-targeting prodrug overcomes tumor resistance to vascular disrupting agents
    Authors: 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
    J. Clin. Invest., 2017;0(0):.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Enzyme Assay
  8. Circulating fibroblast activation protein activity and antigen levels correlate strongly when measured in liver disease and coronary heart disease
    Authors: S Uitte de W, FM Keane, DG Bowen, JJMC Malfliet, HE Zhang, B Maneck, GW McCaughan, FWG Leebeek, DC Rijken, MD Gorrell
    PLoS ONE, 2017;12(6):e0178987.
    Species: Human
    Sample Types: Plasma
    Applications: ELISA (Standard)
  9. Tumor-targeting efficacy of a BF211 prodrug through hydrolysis by fibroblast activation protein-?
    Authors: XP Chai, GL Sun, YF Fang, LH Hu, X Liu, XW Zhang
    Acta Pharmacol. Sin., 2017;0(0):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  10. 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.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Western Blot
  11. Matrilysin/matrix metalloproteinase-7(MMP7) cleavage of perlecan/HSPG2 creates a molecular switch to alter prostate cancer cell behavior.
    Authors: Grindel B, Martinez J, Pennington C, Muldoon M, Stave J, Chung L, Farach-Carson M
    Matrix Biol, 2014;36(0):64-76.
    Species: Human
    Sample Types: Tissue Homogenates
    Applications: Bioassay
  12. Quantitation of fibroblast activation protein (FAP)-specific protease activity in mouse, baboon and human fluids and organs.
    Authors: Keane F, Yao T, Seelk S, Gall M, Chowdhury S, Poplawski S, Lai J, Li Y, Wu W, Farrell P, Vieira de Ribeiro A, Osborne B, Yu D, Seth D, Rahman K, Haber P, Topaloglu A, Wang C, Thomson S, Hennessy A, Prins J, Twigg S, McLennan S, McCaughan G, Bachovchin W, Gorrell M
    FEBS Open Bio, 2013;4(0):43-54.
    Species: N/A
    Sample Types: Recombinant Protein
    Applications: Standard
  13. Radioimmunotherapy of fibroblast activation protein positive tumors by rapidly internalizing antibodies.
    Authors: Fischer E, Chaitanya K, Wuest T, Wadle A, Scott A, van den Broek M, Schibli R, Bauer S, Renner C
    Clin Cancer Res, 2012;18(22):6208-18.
    Species: N/A
    Sample Types: N/A
    Applications: Binding Assay
  14. Neuropeptide Y, B-type natriuretic peptide, substance P and peptide YY are novel substrates of fibroblast activation protein-alpha.
    Authors: Keane FM, Nadvi NA, Yao TW, Gorrell MD
    FEBS J., 2011;278(8):1316-32.
    Species: Human
    Sample Types: Peptide
    Applications: Bioassay

FAQs

  1. Does your Recombinant Human FAP Protein, CF (Catalog # 3715-SE) have a tag?

    • 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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