Recombinant Mouse FAP Protein, CF (2024)

Catalog #: 8647-SE
2 Citations Datasheet / COA / SDS

Analyzed by SEC-MALS

2 Images

All Fibroblast Activation Protein alpha/FAP Proteins and Enzymes

Product Details

Citations (2)

FAQs

Reviews

Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

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

Product Datasheets

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8647-SE

Product Datasheet

COA

SDS

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

Dilute rmFAP to 0.2µg/mL in Assay Buffer.
Dilute Substrate to 100µM in Assay Buffer.
Load 50µL of 0.2µg/mL of rmFAP into a plate, and start the reaction by adding 50µL of 100µM Substrate. Include a Substrate Blank containing 50µL of Assay Buffer and 50 µL of Substrate.
Read at excitation and emission wavelengths of 380nm and 460nm (top read), respectively, in kinetic mode for 5minutes.
Calculate specific activity:

Specific Activity (pmol/min/µg) =	Adjusted V_max* (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

SEC-MALS View Larger

Recombinant Mouse FAP (Catalog # 8647-SE) has a molecular weight (MW) of 173.8 kDa as analyzed by SEC-MALS, suggesting that this protein is a hom*odimer. MW may differ from predicted MW due to post-translational modifications (PTMs) present (i.e. Glycosylation).

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).

Reconstitution Calculator

Background: Fibroblast Activation Protein alpha/FAP

FAP (also known as seprase) is a 95kDa TypeII transmembrane serine protease that is structurally related to dipeptidyl peptidaseIV (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 5aa 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

Zi, F. et al. (2015) Mol. Med. Rep. 11:3203.
Pineiro-Sanchez, M.L. et al. (1997) J. Biol. Chem. 272:7595.
Niedermeyer, J. et al. (1997) Int. J. Cancer 71:383.
Scanlan, M.J. et al. (1994) Proc. Natl. Acad. Sci. USA 91:5657.
Park, J.E. et al. (1999) J. Biol. Chem. 274:36505.
Rettig, W.J. et al. (1988) Proc. Natl. Acad. Sci. USA 85:3110.
Bauer, S. et al. (2006) Arthritis Res. 8:R171.
Aertgeerts, K. et al. (2005) J. Biol. Chem. 280:19441.
Keane, F.M. et al. (2011) FEBS J. 278:1316.
Ghersi, G. et al. (2006) Cancer Res. 66:4652.
Ghersi, G. et al. (2002) J. Biol. Chem. 277:29231.
Cheng, J.D. et al. (2005) Mol. Cancer Ther. 4:351.
Cheng, J.D. et al. (2002) Cancer Res. 62:4767.
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

Related Research Areas

Cancer Biomarkers
DPPIV Family
Other Putative Cancer Biomarkers
Search Bioprocessing Enzymes
Search Immunotherapy-related Enzymes
Serine Proteases and Regulators

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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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, 2017-07-27;12(0):5359-5372.
Species: Mouse
Sample Types: Whole Cells
Applications: Bioassay
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., 2015-12-09;291(15):8070-89.
Applications: Bioassay

FAQs

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 datasheetfor sequence information.

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