Anti-HMGB2 antibody [EPR6302] - BSA and Azide free

• WB

Lab

Western blot - Anti-HMGB2 antibody [EPR6302] - BSA and Azide free (AB248543)

This data was developed using the same antibody clone in a different buffer formulation (ab133540).

Lanes 1- 2 : Merged signal (red and green). Green - ab133540 observed at 24 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab133540 was shown to react with HMGB2 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266358 (knockout cell lysate ab257156) was used. Wild-type HEK-293T and HMGB2 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. ab133540 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 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:

Western blot - Anti-HMGB2 antibody [EPR6302] (<a href='/en-us/products/primary-antibodies/hmgb2-antibody-epr6302-ab133540'>ab133540</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

HMGB2 knockout HEK-293T cell lysate at 20 µg

Predicted band size: 24 kDa

Observed band size: 24 kDa

false

• WB

Lab

Western blot - Anti-HMGB2 antibody [EPR6302] - BSA and Azide free (AB248543)

This data was developed using the same antibody clone in a different buffer formulation (ab133540).

Lanes 1 - 4 : Merged signal (red and green). Green - ab133540 observed at 24 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab133540 was shown to recognize HMGB2 in wild-type HAP1 cells as signal was lost at the expected MW in HMGB2 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and HMGB2 knockout samples were subjected to SDS-PAGE. ab133540 and ab9484 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-HMGB2 antibody [EPR6302] - BSA and Azide free (ab248543) at 1/1000 dilution

Lane 1:

Wild-type HAP1 whole cell lysate at 20 µg

Lane 2:

HMGB2 knockout HAP1 whole cell lysate at 20 µg

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

K562 cell lysate at 20 µg

Predicted band size: 24 kDa

false