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

ab154141 was shown to react with FUS in wild-type HeLa cells in Western blot with loss of signal observed in a FUS knockout cell line. Wild-type HeLa and FUS knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab154141 overnight at 4 °C at a 1/1000 dilution. Blots were incubated with secondary antibodies at 1/5000 before imaging. These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.

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.

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.

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