Detects human Cyclophilin A in direct ELISAs and Western blots. Detects mouse and rat Cyclophilin in Western blots. In direct ELISAs, approximately 50% cross-reactivity
with recombinant human (rh) Cyclophilin E, less than 15% cross-reactivity with
rhCyclophilin F, and less than 5% cross-reactivity with rhCyclophilin A-like 4
Monoclonal Rat IgG2A Clone # 817815
Protein A or G purified from hybridoma culture supernatant
E. coli-derived recombinant human Cyclophilin A Met1-Glu165 Accession # P62937
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Detection of Human, Mouse, and Rat Cyclophilin A by Western Blot. Western blot shows lysates of MOLT‑4 human acute lymphoblastic leukemia cell line, HeLa human cervical epithelial carcinoma cell line, C2C12 mouse myoblast cell line, Nb2‑11 rat lymphoma cell line, and BaF3 mouse pro-B cell line. PVDF membrane was probed with 0.1 µg/mL of Rat Anti-Human Cyclophilin A Monoclonal Antibody (Catalog # MAB3589) followed by HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005). 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 Rat Anti-Human Cyclophilin A Monoclonal Antibody (Catalog # MAB3589) at 10 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 nuclei and cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Preparation and Storage
Sterile PBS to a final concentration of 0.5 mg/mL.
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.
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