- PA1-025 - Invitrogen Antibodies PPIA antibody

Submitted references KIF5B and Nup358 Cooperatively Mediate the Nuclear Import of HIV-1 during Infection.

Dharan A, Talley S, Tripathi A, Mamede JI, Majetschak M, Hope TJ, Campbell EM
PLoS pathogens 2016 Jun;12(6):e1005700

Roles of Capsid-Interacting Host Factors in Multimodal Inhibition of HIV-1 by PF74.

Saito A, Ferhadian D, Sowd GA, Serrao E, Shi J, Halambage UD, Teng S, Soto J, Siddiqui MA, Engelman AN, Aiken C, Yamashita M
Journal of virology 2016 Jun 15;90(12):5808-5823

In vivo functions of CPSF6 for HIV-1 as revealed by HIV-1 capsid evolution in HLA-B27-positive subjects.

Henning MS, Dubose BN, Burse MJ, Aiken C, Yamashita M
PLoS pathogens 2014 Jan;10(1):e1003868

Multiple sites in the N-terminal half of simian immunodeficiency virus capsid protein contribute to evasion from rhesus monkey TRIM5α-mediated restriction.

Kono K, Song H, Yokoyama M, Sato H, Shioda T, Nakayama EE
Retrovirology 2010 Sep 8;7:72

Evidence that mono-ADP-ribosylation of CtBP1/BARS regulates lipid storage.

Bartz R, Seemann J, Zehmer JK, Serrero G, Chapman KD, Anderson RG, Liu P
Molecular biology of the cell 2007 Aug;18(8):3015-25

Proteomic profiling identifies cyclooxygenase-2-independent global proteomic changes by celecoxib in colorectal cancer cells.

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 2006 Sep;15(9):1598-606

Cell surface expression of CD147/EMMPRIN is regulated by cyclophilin 60.

Pushkarsky T, Yurchenko V, Vanpouille C, Brichacek B, Vaisman I, Hatakeyama S, Nakayama KI, Sherry B, Bukrinsky MI
The Journal of biological chemistry 2005 Jul 29;280(30):27866-71

Cyclophilin interactions with incoming human immunodeficiency virus type 1 capsids with opposing effects on infectivity in human cells.

Hatziioannou T, Perez-Caballero D, Cowan S, Bieniasz PD
Journal of virology 2005 Jan;79(1):176-83

The characterization and hormonal regulation of kidney androgen-regulated protein (Kap)-luciferase transgenic mice.

Malstrom SE, Tornavaca O, Meseguer A, Purchio AF, West DB
Toxicological sciences : an official journal of the Society of Toxicology 2004 Jun;79(2):266-77

Redistribution of cyclophilin A to viral factories during vaccinia virus infection and its incorporation into mature particles.

Castro AP, Carvalho TM, Moussatché N, Damaso CR
Journal of virology 2003 Aug;77(16):9052-68

Cyclophilin A peptidyl-prolyl isomerase activity promotes ZPR1 nuclear export.

Ansari H, Greco G, Luban J
Molecular and cellular biology 2002 Oct;22(20):6993-7003

Cholesteryl ester is transported from caveolae to internal membranes as part of a caveolin-annexin II lipid-protein complex.

Uittenbogaard A, Everson WV, Matveev SV, Smart EJ
The Journal of biological chemistry 2002 Feb 15;277(7):4925-31

Native recombinant cyclophilins A, B, and C degrade DNA independently of peptidylprolyl cis-trans-isomerase activity. Potential roles of cyclophilins in apoptosis.

Montague JW, Hughes FM Jr, Cidlowski JA
The Journal of biological chemistry 1997 Mar 7;272(10):6677-84

A calcium-dependent nuclease from apoptotic rat thymocytes is homologous with cyclophilin. Recombinant cyclophilins A, B, and C have nuclease activity.

Montague JW, Gaido ML, Frye C, Cidlowski JA
The Journal of biological chemistry 1994 Jul 22;269(29):18877-80

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot detection of 0.5 µg recombinant human CyPA using Product # PA1-025.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of Cyclophilin A was performed by loading 25 µg of Hela (lane 1), K562 (lane 2), HepG2 (lane 3) and NIH-3T3 (lane 4) cell lysates onto an SDS polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked at 4ºC overnight. The membrane was probed with a Cyclophilin A polyclonal antibody (Product # PA1-025) at a dilution of 1:500 overnight at 4°C, washed in TBST, and probed with an HRP-conjugated secondary antibody for 1 hr at room temperature in the dark. Chemiluminescent detection was performed using Pierce ECL Plus Western Blotting Substrate (Product # 32132). Results show a band at ~18 kDa.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on whole cell extracts and tissue lysates of NIH/3T3 (Lane 1), HeLa (Lane 2), Mouse Lung (Lane 3), Mouse Colon (Lane 4) and Mouse Stomach (Lane 5). The blots were probed with Anti- Cyclophilin A Rabbit Polyclonal Antibody (Product # PA1-025, 1:100- 1:1000 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conj µgate (Product # A27036, 0.4 µg/mL, 1:2500 dilution. A ~ 18 kDa band corresponding to Cyclophilin A was observed across cell lines and tissue lysates tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-Cyclophilin A Polyclonal Antibody(Product # PA1-025) and a 18kDa band corresponding to Cyclophilin A was observed across cell lines tested. Whole cell extracts (30 µg lysate) of A549 (Lane 1), U-87 MG (Lane 2) and Jurkat (Lane 3) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody ( 1:3000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Cyclophilin A (green) showing staining in the in the cytoplasm of A431 cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a Cyclophilin A monoclonal antibody (Product # PA1-025) in 3% BSA-PBS at a dilution of 1:200 and incubated overnight at 4ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Cyclophilin A (green) showing staining in the in the cytoplasm of Hela cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a Cyclophilin A polyclonal antibody (Product # PA1-025) in 3% BSA-PBS at a dilution of 1:200 and incubated overnight at 4ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: 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 (Product # PA1-025) at 1:250 dilution 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 conjugate (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 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.