Detection of Human, Mouse, and Rat Cyclophilin A by Western Blot.
Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line, C2C12 mouse myoblast cell line, and Nb2‑11 rat lymphoma cell line. PVDF membrane was probed with 0.1 µg/mL of Goat Anti-Human Cyclophilin A Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3589) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for Cyclophilin A at approximately 18 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Cyclophilin A in PANC-1 Human Cell Line.
Cyclophilin A was detected in immersion fixed PANC-1 human pancreatic carcinoma cell line using Goat Anti-Human/Mouse/Rat Cyclophilin A Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3589) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to nuclei and cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Detection of Human and Mouse Cyclophilin A by Simple WesternTM.
Simple Western lane view shows lysates of HeLa human cervical epithelial carcinoma cell line and C2C12 mouse myoblast cell line, loaded at 0.2 mg/mL. A specific band was detected for Cyclophilin A at approximately 25 kDa (as indicated) using 5 µg/mL of Goat Anti-Human/Mouse/Rat Cyclophilin A Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3589) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
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. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
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: Cyclophilin A
Cyclophilin A, also called Peptidyl-prolyl Isomerase A, PPIA, CYPA, and CYPH, was originally characterized for its ability to catalyze the transition between cis- and trans- proline residues critical for proper folding of proteins (1). Cyclophilin is also incorporated into many viruses, including HIV-1, where it has been speculated to be involved in functions such as viral assembly and infectivity (2). The immunosuppressive activity of cyclosporins has been correlated with their ability to form complexes with cyclophilins that inhibit calcineurin phosphatase activity (3) and prevent incorporation of cyclophilin into viral particles (4). The cyclosporin/cyclophilin complex selectively binds and inactivates calcineurin (3, 5), making it a useful inhibitor for studying calcineurin activity.
Hamilton, G.S. and J.P. Steiner (1998) J. Med. Chem. 41:5119.
Cantin, R. et al. (2005) J. Virology 79:6577.
Liu, J. et al. (1992) Biochemistry 31:3896.
Wiegers K. and H.G. Krausslich (2002) Virology 294:289.
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.
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