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Immunofluorescence analysis of Cyclophilin A was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Cyclophilin A Rabbit Polyclonal Antibody (PA1025) at 2 µg/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conj µgate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Hamster, Human, Mouse|
|Published species reactivity||Rat, Human, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Purified recombinant human CyPA.|
|Purification||Ammonium sulfate precipitation|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Western Blot (WB)||1:100-1:1000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Western Blot (WB)||See 8 publications below|
PA1-025 detects recombinant human cyclophilin A (CyPA), but does not detect endogenous levels of CyPA. PA1-025 also detects the CyPA protein from CHO cells.
PA1-025 has been successfully used in Western blot and ICC/IF procedures. By Western blot, this antibody detects a prominent ~18 kDa protein representing recombinant human CyPA.
The PA1-025 immunogen is recombinant human CyPA expressed in E. coli.
Immunophilins are a family of soluble cytosolic receptors capable of binding to one of two major immunosuppressant agents - cyclosporin A (CsA) or FK506. Proteins that bind FK506 are termed FK506 Binding Proteins (FKBPs) and those that bind cyclosporin A are called cyclophilins (CyP).
Both CyP:CsA and FKBP:FK506 complexes have been shown to inhibit calcineurin, a calcium and calmodulin dependent protein phosphatase which has been implicated as an important signaling enzyme in T-cell activation, providing a possible mechanism of immunosuppression by CsA and FK506. Immunophilins function as peptidyl prolyl cis-trans-isomerases (PPIase) whose activity is inhibited by their respective immunosuppressant compounds. As PPIase's, immunophilins accelerate folding of some proteins both in vivo and in vitro by catalyzing slow steps in the initial folding and rearrangement of proline containing proteins.
Within the cyclophilin family, there are several different proteins which show a high degree of homology including CyPA, CyPB and CyPC. CyPA, also termed CyP-18, is the most abundant and ubiquitous cyclophilin found in all vertebrate tissues and is present in T-cells.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Multiple sites in the N-terminal half of simian immunodeficiency virus capsid protein contribute to evasion from rhesus monkey TRIM5α-mediated restriction.
PA1-025 was used in western blot to investigate the mechanism for the evasion of Rh TRIM5alpha restriction by simian immunodeficiency viru core protein
|Kono K,Song H,Yokoyama M,Sato H,Shioda T,Nakayama EE||Retrovirology (7:null)||2010|
Proteomic profiling identifies cyclooxygenase-2-independent global proteomic changes by celecoxib in colorectal cancer cells.
PA1-025 was used in western blot to find new celecoxib-mediated markers in the COX-2-deficient colorectal cancer cell line
|Lou J,Fatima N,Xiao Z,Stauffer S,Smythers G,Greenwald P,Ali IU||Cancer epidemiology, biomarkers & prevention : a publication of the American Association for Cancer Research, cosponsored by the American Society of Preventive Oncology (15:1598)||2006|
Cell surface expression of CD147/EMMPRIN is regulated by cyclophilin 60.
PA1-025 was used in western blot to demonstrate the role of Cyp60 in the CD147 translocation to the cell surface.
|Pushkarsky T,Yurchenko V,Vanpouille C,Brichacek B,Vaisman I,Hatakeyama S,Nakayama KI,Sherry B,Bukrinsky MI||The Journal of biological chemistry (280:27866)||2005|
Cyclophilin interactions with incoming human immunodeficiency virus type 1 capsids with opposing effects on infectivity in human cells.
PA1-025 was used in western blot to investigate the role of CypA-CA interaction in regulating HIV-1 replication and infectivity in target human cells.
|Hatziioannou T,Perez-Caballero D,Cowan S,Bieniasz PD||Journal of virology (79:176)||2005|
The characterization and hormonal regulation of kidney androgen-regulated protein (Kap)-luciferase transgenic mice.
PA1-025 was used in western blot to study the expression of kidney androgen regulated protein (Kap) in Kap-luc transgenic mice
|Malstrom SE,Tornavaca O,Meseguer A,Purchio AF,West DB||Toxicological sciences : an official journal of the Society of Toxicology (79:266)||2004|
Cyclophilin A peptidyl-prolyl isomerase activity promotes ZPR1 nuclear export.
PA1-025 was used in western blot to investigate the role for Cpr1p in Zpr1p nuclear export and its functional PPIase activity.
|Ansari H,Greco G,Luban J||Molecular and cellular biology (22:6993)||2002|
Native recombinant cyclophilins A, B, and C degrade DNA independently of peptidylprolyl cis-trans-isomerase activity. Potential roles of cyclophilins in apoptosis.
PA1-025 was used in western blot to investigate the DNA degradation by three cyclophilins isoforms
|Montague JW,Hughes FM,Cidlowski JA||The Journal of biological chemistry (272:6677)||1997|
A calcium-dependent nuclease from apoptotic rat thymocytes is homologous with cyclophilin. Recombinant cyclophilins A, B, and C have nuclease activity.
PA1-025 was used in western blot to investigate the functional properties of NUC18 compared to cyclophilin
|Montague JW,Gaido ML,Frye C,Cidlowski JA||The Journal of biological chemistry (269:18877)||1994|
cyclophilin A; cyclosporin A-binding protein; CyP A; EC 188.8.131.52; epididymis secretory sperm binding protein Li 69p; peptidyl-prolyl cis-trans isomerase A; peptidylprolyl isomerase A (cyclophilin A); PPIase A; rotamase A; SP18; T cell cyclophilin
2700098C05; Cphn; CyP-18; CYPA; CYPH; HEL-S-69p; PPIA