All lanes : Anti-Cyclophilin B antibody [CL3901] (ab236760) at 1/1000 dilution

Lane 1 : Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : PPIB knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 3 : U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) whole cell lysate
Lane 4 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

Predicted band size: 24 kDa
Observed band size: 24 kDa

Lanes 1-4: Merged signal (red and green). Green - ab236760 observed at 24 kDa. Red - loading control ab8245 observed at 36 kDa.

 ab236760 Anti-Cyclophilin B antibody [CL3901]  was shown to specifically react with Cyclophilin B in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261746 (knockout cell lysate ab257037) was used. Wild-type and Cyclophilin B knockout samples were subjected to SDS-PAGE. ab236760 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

Formalin-fixed, paraffin-embedded human stomach tissue stained for Cyclophilin B with ab236760 at a 1/5000 dilution in immunohistochemical analysis.

All lanes : Anti-Cyclophilin B antibody [CL3901] (ab236760) at 1/5000 dilution

Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : PPIB knockout HAP1 whole cell lysate
Lane 3 : Jurkat whole cell lysate
Lane 4 : U87-MG whole cell lysate

Lysates/proteins at 20 µg per lane.

Predicted band size: 24 kDa

Lanes 1 - 4: Merged signal (red and green). Green - ab236760 observed at 24 kDa. Red - loading control, ab181602, observed at 37 kDa.

ab236760 was shown to specifically react with PPIB in wild-type HAP1 cells as signal was lost in PPIB knockout cells. Wild-type and PPIB knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. Ab236760 and ab181602 (Rabbit anti-GAPDH loading control) were incubated overnight at 4°C at 1/5000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye^® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes : Anti-Cyclophilin B antibody [CL3901] (ab236760) at 1 µg/ml

Lane 1 : Control siRNA transfected U-251 MG (human brain glioma cell line) cell lysate
Lane 2 : Cyclophilin B siRNA probe 1 transfected U-251 MG (human brain glioma cell line) cell lysate
Lane 3 : Cyclophilin B siRNA probe 2 transfected U-251 MG (human brain glioma cell line) cell lysate

Predicted band size: 24 kDa

All lanes : Anti-Cyclophilin B antibody [CL3901] (ab236760) at 1 µg/ml

Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) cell lysate
Lane 2 : HEK-293 (human epithelial cell line from embryonic kidney) cell lysate
Lane 3 : A431 (human epidermoid carcinoma cell line) cell lysate
Lane 4 : HepG2 (human liver hepatocellular carcinoma cell line) cell lysate
Lane 5 : NIH/3T3 (mouse embryo fibroblast cell line) cell lysate
Lane 6 : NBT II (rat cell line) cell lysate

Predicted band size: 24 kDa

Formalin-fixed, paraffin-embedded human placenta tissue stained for Cyclophilin B with ab236760 at a 1/5000 dilution in immunohistochemical analysis.

Formalin-fixed, paraffin-embedded human prostate tissue stained for Cyclophilin B with ab236760 at a 1/5000 dilution in immunohistochemical analysis.

Formalin-fixed, paraffin-embedded human skeletal muscle tissue stained for Cyclophilin B with ab236760 at a 1/5000 dilution in immunohistochemical analysis.