Cyclophilin A Antibody [Unconjugated]

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.

Simple Western shows lysates of Exosome Standards (HT‑29) (NBP3-11685), Exosome Standards (PC‑3) (NBP2-49856) and MOLT‑4 human acute lymphoblastic leukemia cell line, loaded at 0.5 mg/ml. A specific band was detected for Cyclophilin A at approximately 23 kDa (as indicated) using 50 µg/mL of Goat Anti-Human/Mouse/Rat Cyclophilin A Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3589). This experiment was conducted under reducing conditions and using the 2-40kDa separation system.

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 Northern-Lights™ 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.

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.

• Reactivity: Hu, Mu, Rt
• Applications: WB, Simple Western, ICC/IF
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

Cyclophilin A Antibody [Unconjugated] Summary

• Immunogen: E. coli-derived recombinant human Cyclophilin A
  Met1-Glu165
  Accession # P62937
• Specificity: Detects human, mouse, and rat Cyclophilin A in Western blots.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: PPIA
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 5-15 ug/mL
  • Simple Western 5 ug/mL
  • Western Blot 0.1 ug/mL
• Application Notes: In Simple Western only 10-15 uL of the recommended dilution is used per data point.

Packaging, Storage & Formulations

• 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.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details.
  *Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Cyclophilin A Antibody [Unconjugated]

• Cyclophilin A
• Cyclosporin A-binding protein
• CYPA
• CYPAMGC12404
• CYPH
• EC 5.2.1.8
• MGC117158
• MGC23397
• peptidyl-prolyl cis-trans isomerase A
• peptidylprolyl isomerase A (cyclophilin A)
• PPIA
• PPIase A
• Rotamase A
• Rotamase
• T cell cyclophilin

Background

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.

1. Hamilton, G.S. and J.P. Steiner  (1998) J. Med. Chem. 41:5119.
2. Cantin, R. et al. (2005) J. Virology 79:6577.
3. Liu, J. et al. (1992) Biochemistry 31:3896.
4. Wiegers K. and H.G. Krausslich (2002) Virology 294:289.
5. Liu, J. et al. (1991) Cell 66:807.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Cyclophilin A Antibody (AF3589)(2)

Publications using AF3589

• Xuefeng Deng, Wei Zou, Min Xiong, Zekun Wang, John F. Engelhardt, Shui Qing Ye et al. Human Parvovirus Infection of Human Airway Epithelia Induces Pyroptotic Cell Death by Inhibiting Apoptosis Journal of Virology 2017-12-15 [PMID: 29021400]
• Heinzmann D, Bangert A, Muller A, von Ungern-Sternberg S, Emschermann F, Schonberger T, Chatterjee M, Mack A, Klingel K, Kandolf R, Malesevic M, Borst O, Gawaz M, Langer H, Katus H, Fischer G, May A, Kaya Z, Seizer P The Novel Extracellular Cyclophilin A (CyPA) - Inhibitor MM284 Reduces Myocardial Inflammation and Remodeling in a Mouse Model of Troponin I -Induced Myocarditis. PLoS ONE, 2015-04-20;10(4):e0124606. 2015-04-20 [PMID: 25894208] (IHC, Mouse)

Bioinformatics

• Gene Symbol: PPIA
• Uniprot:
  • P62937()