C2C12 mouse myoblast cell line was stained with Rat Anti-Mouse Fibroblast Activation Protein alpha/FAP Monoclonal Antibody (Catalog # MAB9727, filled histogram) or isotype control antibody (Catalog # MAB005, open histogram), followed by Phycoerythrin-conjugated Anti-Rat IgG Secondary Antibody (Catalog # F0105B). View our protocol for Staining Membrane-associated Proteins.

Fibroblast Activation Protein alpha/FAP in C2C12 Mouse Cell Line.

Fibroblast Activation Protein a/FAP was detected in immersion fixed C2C12 mouse myoblast cell line using Rat Anti-Mouse Fibroblast Activation Protein a/FAP Monoclonal Antibody (Catalog # MAB9727) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rat IgG Secondary Antibody (red; Catalog # NL013) and counterstained with DAPI (blue). Specific staining was localized to plasma membrane and cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Detection of Fibroblast Activation Protein alpha/FAP in NIH-3T3 Mouse Embryonic Fibroblast Cell Line by Flow Cytometry.

Used 3uL for 15mins at 4°C on 3T3 cells transduced to over express murine FAP. Image from a verified customer review.

Applications for Mouse Fibroblast Activation Protein alpha/FAP Antibody

Application

Recommended Usage

ELISA

This antibody functions as an ELISA capture antibody when paired with Rat Anti-Mouse Fibroblast Activation Protein alpha/FAP Monoclonal Antibody (Catalog # MAB97271).

This product is intended for assay development on various assay platforms requiring antibody pairs.

Flow Cytometry

0.25 µg/10⁶ cells
Sample: C2C12 mouse myoblast cell line

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed C2C12 mouse myoblast cell line

Reviewed Applications

Read 1 review rated 5 using MAB9727 in the following applications:

Flow Cytometry (1 Review)

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

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

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.

Alternate Names

FAP, Seprase

Entrez Gene IDs

2191 (Human); 14089 (Mouse); 102134935 (Cynomolgus Monkey)

Gene Symbol

FAP

UniProt

P97321

Additional Fibroblast Activation Protein alpha/FAP Products

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Product Specific Notices for Mouse Fibroblast Activation Protein alpha/FAP Antibody

For research use only

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Flow Cytometry

Mouse Fibroblast Activation Protein alpha/FAP Antibody

R&D Systems, part of Bio-Techne | Catalog # MAB9727

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Scientific Data Images for Mouse Fibroblast Activation Protein alpha/FAP Antibody

Detection of Fibroblast Activation Protein alpha/FAP in C2C12 Mouse Cell Line by Flow Cytometry.