Lane 1: Wild-type MCF7 cell lysate (20µg)

Lane 2: BMI1 knockout MCF7 cell lysate (20µg)
Lane 3: A431 cell lysate (20µg)
Lane 4: HEK-293 cell lysate (20µg)
Lanes 1- 4: Merged signal (red and green). Green - ab269678 observed at 36 kDa. Red - loading control ab52866 observed at 50 kDa.
ab269678 Mouse monoclonal [BMI1/2823] to Bmi1 was shown to specifically react with Bmi1 in wild-type MCF7 cells in western blot. Loss of signal was observed when knockout cell line ab262319 (knockout cell lysate ab256851) was used. Wild-type and Bmi1 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab269678 and Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab52866) were incubated overnight at 4^°C at 2 dilution and 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.

Lane 1: Wild-type MCF7 cell lysate (20µg)

Lane 2: BMI1 knockout MCF7 cell lysate (20µg)
Lane 3: A431 cell lysate (20µg)
Lanes 1- 3: Merged signal (red and green). Green - ab126783 observed at 37 kDa. Red - loading control ab7291 observed at 50 kDa.
ab126783 Rabbit monoclonal [EPR3745(2)] to Bmi1 was shown to specifically react with Bmi1 in wild-type MCF7 cells in western blot. Loss of signal was observed when knockout cell line ab262319 (knockout cell lysate ab256851) was used. Wild-type and Bmi1 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab126783 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4^°C at 1 in 10000 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.

Allele-1: 1 bp deletion in exon2

 

Allele-2: 1 bp insertion in exon2