All lanes : Anti-beta 2 Microglobulin antibody [EP2978Y] (ab75853) at 1/1000 dilution

Lane 1 : Wild-type HepG2 cell lysate
Lane 2 : B2M knockout HepG2 cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : Jurkat 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: 14 kDa
Observed band size: 14 kDa

Lanes 1-4: Merged signal (red and green). Green - ab75853 observed at 14 kDa. Red - loading control ab8245 observed at 36 kDa.

 ab75853 Anti-beta 2 Microglobulin antibody [EP2978Y] was shown to specifically react with beta 2 Microglobulin in wild-type HepG2 cells. Loss of signal was observed when knockout cell line ab262325 (knockout cell lysate ab256846) was used. Wild-type and beta 2 Microglobulin knockout samples were subjected to SDS-PAGE. ab75853 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.

All lanes : Anti-beta 2 Microglobulin antibody [EPR21752-214] (ab218230) at 1/500 dilution

Lane 1 : Wild-type HepG2 cell lysate
Lane 2 : B2M knockout HepG2 cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : Jurkat 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: 14 kDa
Observed band size: 14 kDa

Lanes 1-4: Merged signal (red and green). Green - ab218230 observed at 14 kDa. Red - loading control ab8245 observed at 36 kDa.

 ab218230 Anti-beta 2 Microglobulin antibody [EPR21752-214]  was shown to specifically react with beta 2 Microglobulin in wild-type HepG2 cells. Loss of signal was observed when knockout cell line ab262325 (knockout cell lysate ab256846) was used. Wild-type and beta 2 Microglobulin knockout samples were subjected to SDS-PAGE. ab218230 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 500 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 insertion in exon1

 

Allele-2: Insertion of the selection cassette in exon 1.