Recombinant Human Cyclophilin A Protein, CF

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R&D Systems Recombinant Proteins and Enzymes
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Citations (9)

Recombinant Human Cyclophilin A Protein, CF Summary

Product Specifications

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
Measured by its ability to inhibit calcineurin phosphatase activity in the presence of Cyclosporin A. The IC50 for inhibition of calcineurin activity is <700 nM, as measured under the described conditions.
E. coli-derived human Cyclophilin A protein
Accession #
N-terminal Sequence
Predicted Molecular Mass
18 kDa
17 kDa, reducing conditions

Product Datasheets

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Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.


Formulation Supplied as a 0.2 μm filtered solution in MES and NaCl.
Shipping The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.

Assay Procedure

  • Assay Buffer (AB): 20 mM Tris, 10 mM MgCl2, 0.1 mM CaCl2, 1 mg/mL BSA, pH 7.5
  • Recombinant Human Cyclophilin A (rhCyclophilin A) (MW: 18 kDa) (Catalog # 3589-CAB)
  • Recombinant Human Calcineurin (rhCalcineurin) (Catalog # 3160-CA)
  • Substrate: Serine/Threonine Phosphatase Substrate I (Catalog # ES012), 10 mM stock in deionized water
  • Calmodulin (Sigma, Catalog # P1431), 0.168 mg/mL stock in Assay Buffer
  • EDTA (Sigma, Catalog # E6758), 0.1 M stock in deionized water (pH to 8.0)
  • Cyclosporin A (Sigma, Catalog # C1832), 1 mM stock in DMSO
  • Incubator at 37 °C able to shake microplate
  • Malachite Green Phosphate Detection Kit (Catalog # DY996)
  • 96-well Clear Plate (Catalog # DY990)
  • Plate Sealers (Corning, Catalog # 3095)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Dilute Calcineurin to 1.17 µg/mL, Calmodulin to 16.8 µg/mL (1 µM), and Cyclosporin A to 30 µM in Assay Buffer (AB).
  2. Dilute Cyclophilin A in AB to the following concentrations: neat, 10000, 5000, 2500, 1250, 625, 313, 156, 78, 39, and 20 nM.
  3. Add the following to the wells of a plate (add largest volume last to mix all together):
    5 µL of Cyclophilin A dilutions
    5 µL of Calmodulin
    5 µL of Cyclosporin A
    25 µL of Calcineurin
    Pos. Control (prepare two)        
    5 µL of assay buffer
    5 µL of Calmodulin
    5 µL of Cyclosporin A
    25 µL of Calcineurin
    Neg. Control (optional, prepare one)
    5 µL of EDTA
    5 µL of Calmodulin
    5 µL of Cyclosporin A
    25 µL of Calcineurin
  4. Seal plate, tap to mix and incubate plate at room temperature for 1 hr.
  5. After incubation remove seal and add 10 µL of 1 mM Substrate.
  6. Reseal and incubate at 37 °C with shaking for 1 hr.
  7. During second incubation, prepare kit standard curve. Prepare serial dilutions in AB at the following conc.: 100, 50, 25, 12.5, 6.25, 3.13, and 1.57 µM.
  8. Add 50 µL of the phos. curve to the plate, in duplicate, including a 0 mM phos. point (50 µL AB).
  9. To all wells add 10 µL of kit reagent A, tap to mix, and incubate at RT for 10 min.
  10. To all wells add 10 µL of kit reagent B, tap to mix, and incubate at RT for 20 min.
  11. Read at 620 nm.
  12. Determine the 50% inhibiting conc. (IC50) of rhCyclophilin A by plotting conc. (nM) vs. Phos. released (nmol) with 4-PL fitting.
  13. The specific activity of rhCalcineurin at each point can be determined using the following eqn. (if needed):

     Specific Activity (nmol/min/mg) =

Phosphate released* (nmol)
Inc. time (min) x amount of enzyme (mg)

     *Derived from the phosphate standard curve using linear or 4-parameter fitting and adjusted for Negative Control.

Per Well:
  • Phos. standard: 5.0, 2.5, 1.25, 0.625, 0.313, 0.156, 0.078, and 0 nmol
  • rhCyclophilin A: nt/14, 714, 357, 179, 89, 45, 22, 11, 5.6, 2.8, 1.4, 0.0 nM 
  • rhCalcineurin: 0.00002925 mg
Reconstitution Calculator

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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.

  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.
Entrez Gene IDs
5478 (Human); 268373 (Mouse); 25518 (Rat)
Alternate Names
Cyclophilin A; Cyclosporin A-binding protein; CYPA; CYPAMGC12404; CYPH; EC; MGC117158; MGC23397; peptidyl-prolyl cis-trans isomerase A; peptidylprolyl isomerase A (cyclophilin A); PPIA; PPIase A; Rotamase A; Rotamase; T cell cyclophilin

Citations for Recombinant Human Cyclophilin A Protein, CF

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.

9 Citations: Showing 1 - 9
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  1. p52 signaling promotes cellular senescence
    Authors: GM Bernal, L Wu, DJ Voce, RR Weichselba, B Yamini
    Cell & bioscience, 2022;12(1):43.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  2. Cyclophilin A Is Not Acetylated at Lysine-82 and Lysine-125 in Resting and Stimulated Platelets
    Authors: A Rosa, E Butt, CP Hopper, S Loroch, M Bender, H Schulze, A Sickmann, S Vorlova, P Seizer, D Heinzmann, A Zernecke
    International Journal of Molecular Sciences, 2022;23(3):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  3. Cerebral Apolipoprotein D Exits the Brain and Accumulates in Peripheral Tissues
    Authors: F Desmarais, V Hervé, KF Bergeron, G Ravaut, M Perrotte, G Fyfe-Desma, E Rassart, C Ramassamy, C Mounier
    International Journal of Molecular Sciences, 2021;22(8):.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Bioassay
  4. APOE4 leads to blood-brain barrier dysfunction predicting cognitive decline
    Authors: A Montagne, DA Nation, AP Sagare, G Barisano, MD Sweeney, A Chakhoyan, M Pachicano, E Joe, AR Nelson, LM D'Orazio, DP Buennagel, MG Harrington, TLS Benzinger, AM Fagan, JM Ringman, LS Schneider, JC Morris, EM Reiman, RJ Caselli, HC Chui, J Tcw, Y Chen, J Pa, PS Conti, M Law, AW Toga, BV Zlokovic
    Nature, 2020;581(7806):71-76.
    Species: Human
    Sample Types: CSF
    Applications: ELISA Standard
  5. MGAT1 and Complex N-Glycans Regulate ERK Signaling During Spermatogenesis
    Authors: B Biswas, F Batista, S Sundaram, P Stanley
    Sci Rep, 2018;8(1):2022.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Bioassay
  6. MGAT1 and Complex N-Glycans Regulate ERK Signaling During Spermatogenesis.
    Authors: Barnali Biswas, Frank Batista, Subha Sundaram, Pamela Stanley
    Scientific Reports, 2018;0(0):2045-2322.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  7. Cyclophilin A enhances macrophage differentiation and lipid uptake in high glucose conditions: a cellular mechanism for accelerated macro vascular disease in diabetes mellitus
    Cardiovasc Diabetol, 2016;15(1):152.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  8. The Novel Extracellular Cyclophilin A (CyPA) - Inhibitor MM284 Reduces Myocardial Inflammation and Remodeling in a Mouse Model of Troponin I -Induced Myocarditis.
    Authors: 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
    PLoS ONE, 2015;10(4):e0124606.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  9. EMMPRIN and its ligand cyclophilin A regulate MT1-MMP, MMP-9 and M-CSF during foam cell formation.
    Authors: Seizer P, Schonberger T, Schott M, Lang MR, Langer HF, Bigalke B, Kramer BF, Borst O, Daub K, Heidenreich O, Schmidt R, Lindemann S, Herouy Y, Gawaz M, May AE
    Atherosclerosis, 2010;209(1):51-7.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay


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