Recombinant Mouse FAP Protein, CF

Catalog # Availability Size / Price Qty
8647-SE-010
Recombinant Mouse FAP Protein Enzyme Activity
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Product Details
Citations (2)
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Recombinant Mouse 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 >2000 pmol/min/μg, as measured under the described conditions.
Source
Mouse myeloma cell line, NS0-derived mouse Fibroblast Activation Protein alpha/FAP protein
Leu26-Asp761
Accession #
N-terminal Sequence
Analysis
Leu26 & Val31
Predicted Molecular Mass
85 kDa
SDS-PAGE
78-90 kDa, reducing conditions

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8647-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.

8647-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.

Assay Procedure

Materials
  • Assay Buffer: 50 mM Tris, 1 M NaCl, 1 mg/mL BSA, pH 7.5
  • Recombinant Mouse Fibroblast Activation Protein alpha /FAP (rmFAP) (Catalog # 8647-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 rmFAP 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 rmFAP 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 # A9891).

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

Scientific Data

Enzyme Activity Recombinant Mouse FAP Protein Enzyme Activity View Larger

Recombinant Mouse FAP (Catalog # 8647-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).

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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, mouse FAP shares 90% and 97% amino acid (aa) sequence identity with human and rat FAP, respectively (3, 4). Alternative splicing of mouse FAP generates isoforms with a 33 aa or 5 aa deletion in the extracellular juxtamembrane region (3). FAP is expressed on reactive stromal fibroblasts in tumor tissue and wound healing and on synoviocytes in rheumatoid arthritis (1, 5-7). It exhibits dipeptidyl peptidase activity with substrate specificity similar to DPPIV, which is specific for N-terminal Xaa-Pro sequences (5, 8). FAP is also an endopeptidase that can degrade Gelatin, Collagens I and IV, Fibronectin, and Laminin (1, 5, 8) as well as several peptide hormones (e.g. Neuropeptide Y, Brain Natriuretic Peptide, Substance P, Peptide YY, and Incretins) (9). The enzymatic activity is dependent on FAP association with DPPIV on the cell surface (5, 8, 10, 11). The matrix-dedgrading activity of FAP contributes to tumor cell migration and invasion (10-13). In addition, FAP can enhance tumor cell growth by limiting the development of anti-tumor immunity (14).

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 Mouse 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.

2 Citations: Showing 1 - 2
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  1. A synthetic urinary probe-coated nanoparticles sensitive to fibroblast activation protein ? for solid tumor diagnosis
    Authors: X Feng, Q Wang, Y Liao, X Zhou, Y Wang, W Liu, G Zhang
    Int J Nanomedicine, 2017;12(0):5359-5372.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Bioassay
  2. Fibroblast Activation Protein (FAP) Accelerates Collagen Degradation and Clearance from Lungs in Mice
    Authors: MH Fan, Q Zhu, HH Li, HJ Ra, S Majumdar, DL Gulick, JA Jerome, DH Madsen, M Christofid, DW Speicher, WW Bachovchin, C Feghali-Bo, E Puré
    J. Biol. Chem., 2016;291(15):8070-89.
    Species: N/A
    Sample Types: Recombinant Protein
    Applications: Bioassay

FAQs

  1. Does this enzyme have a tag?

    • 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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