Detection of Human Fibroblast Activation Protein alpha /FAP by Western Blot.
Western blot shows lysates of WI‑38 human lung fibroblast cell line. PVDF membrane was probed with 0.5 µg/mL of Sheep Anti-Human Fibroblast Activation Protein alpha /FAP Biotinylated Antigen Affinity-purified Polyclonal Antibody (Catalog # BAF3715) followed by Streptavidin-HRP (Catalog # DY998). A specific band was detected for Fibroblast Activation Protein alpha /FAP at approximately 97 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it 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 Type II transmembrane serine protease that is structurally related to dipeptidyl peptidase IV (1). FAP has substrate specificity similar to dipeptidyl peptidase IV, which is specific for N-terminal Xaa-Pro sequences, but FAP is also an endopeptidase able to degrade gelatin and Type I collagen (2). The enzymatically active form of FAP is a dimer (3). FAP has a restricted tissue distribution. It is not detectable in normal tissues or resting fibroblasts, but is highly expressed on reactive stromal fibroblasts in epithelial cancers (4), in granulation tissue during wound healing, and in bone and soft tissue sarcomas (5). Because of its expression patterns and enzymatic activities, FAP is believed to play roles in tumor invasion, tissue remodeling, and wound repair.
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Pineiro-Sanchez, M.L. et al. (1997) J. Biol. Chem. 272:7595.
Garin-Chesa, P. et al. (1990) Proc. Natl. Acad. Sci. USA 87:7235.
Rettig, W.J. et al. (1988) Proc. Natl. Acad. Sci. USA 85:3110.
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We have 1 review tested in 1 species: Mouse.
We have 1 review tested in 1 application: Flow Cytometry.
Flow Cytometry: Human Fibroblast Activation Protein alpha /FAP Biotinylated Antibody [BAF3715].
Tumor cell lyastes
Other Experimental Details
Other Experimental Details
Since the mouse FAP shares ~90% of the human protein sequence, in our hands this antibody works good also mouse cells.We are using it for flow cytometry on primary mouse cells in combination with APC Streptavidin (Biolegend, 405207). The staining works properly with a concentration of 1:20 of FAP Biotinylated Antibody and 1:100 of APC Streptavidin.