All lanes : Anti-HMGB1 antibody (ab77302) at 5 µg/mlLane 1 : HMGB1 transfected 293T cell lysateLane 2 : Non-transfected 293T cell lysate Lysates/proteins at 25 µg per lane.SecondaryGoat Anti-Mouse IgG (H&L)-HRP Conjugate at 1/2500 dilution
Immunoperoxidase of ab77302, at 3µg/ml, staining HMGB1 in formalin-fixed, paraffin-embedded human stomach tissue.
ab77302, at 10µg/ml, staining HMGB1 in Hela cells by Immunofluorescence.
![Overlay histogram showing HeLa cells stained with ab77302 (red line). The cells were fixed with methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab77302, 2µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween 20 used under the same conditions.](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_3/2914_HMGB1-Primary-antibodies-ab77302-1.jpg)
Overlay histogram showing HeLa cells stained with ab77302 (red line). The cells were fixed with methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab77302, 2µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween 20 used under the same conditions.
Immunofluorescence analysis of Human peripheral polymorphonuclear cells, staining HMGB1 with ab77302. Cells were fixed in 4% paraformaldehyde and blocked with 5% rabbit serum. Cells were incubated with primary antibody and an AlexaFluor®488-conjugated anti-mouse IgG was used as the secondary antibody. Nuclei were stained with DAPI.