All lanes : Anti-TLS/FUS antibody [CL0190] (ab154141) at 1/500 dilution

Lane 1 : Wild-type HAP1 cell lysate
Lane 2 : TLS/FUS knockout HAP1 cell lysate
Lane 3 : K562 cell lysate
Lane 4 : HepG2 cell lysate

Lysates/proteins at 20 µg per lane.

Predicted band size: 53 kDa

Lanes 1 - 4: Merged signal (red and green). Green - ab154141 observed at 80 kDa. Red - loading control, ab181602, observed at 37 kDa.

ab154141 was shown to specifically react with TLS/FUS when TLS/FUS knockout samples were used. Wild-type and TLS/FUS knockout samples were subjected to SDS-PAGE. ab154141 and ab181602 (loading control to GAPDH) were diluted at 1/500 and 1/10000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Mouse IgG H&L (IRDye^® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye^® 680RD)preadsorbed (ab216777) secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging.

Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling TLS/FUS with ab154141 showing nuclear (without nucleoli) staining in green. Microtubule are visualized in red.

All lanes : Anti-TLS/FUS antibody [CL0190] (ab154141) at 1/500 dilution

Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : FUS knockout HEK-293T cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 53 kDa
Observed band size: 80 kDa why is the actual band size different from the predicted?

Lanes 1- 2: Merged signal (red and green). Green - ab154141 observed at 80 kDa. Red - Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) observed at 37 kDa.

 ab154141 was shown to react with TLS/FUS in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266587 (knockout cell lysate ab257100) was used. Wild-type HEK-293T and FUS knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab154141 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) overnight at 4°C at a 1 in 500 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye^®800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye^®680RD) preadsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes : Anti-TLS/FUS antibody [CL0190] (ab154141)

Lane 1 : U-251 cells, transfected with control siRNA
Lane 2 : U-251 cells, transfected with target specific siRNA probe #1
Lane 3 : U-251 cells, transfected with target specific siRNA probe #2

Predicted band size: 53 kDa

Downregulation of antibody signal confirms target specificity. Remaining % intensity, relative control lane, is indicated. Anti-GAPDH monoclonal antibody was used as loading control.

Immunohistochemical analysis of Human kidney tissue labeling TLS/FUS with ab154141 at 1/2500 dilution.

Immunohistochemical analysis of Human prostate tissue labeling TLS/FUS with ab154141 at 1/2500 dilution.

Immunohistochemical analysis of Human stomach tissue labeling TLS/FUS with ab154141 at 1/2500 dilution.

Immunohistochemical analysis of Human pancreas tissue labeling TLS/FUS with ab154141 at 1/2500 dilution.