Anti-HMGB1 antibody (ab77302)

  Western blot - HMGB1 antibody (ab77302)

All lanes : Anti-HMGB1 antibody (ab77302) at 5 µg/ml

Lane 1 : HMGB1 transfected 293T cell lysate
Lane 2 : Non-transfected 293T cell lysate

Lysates/proteins at 25 µg per lane.

Secondary
Goat Anti-Mouse IgG (H&L)-HRP Conjugate at 1/2500 dilution

Predicted band size : 25 kDa
Observed band size : 29 kDa (why is the actual band size different from the predicted?)

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - HMGB1 antibody (ab77302)

Immunoperoxidase of ab77302, at 3µg/ml, staining HMGB1 in formalin-fixed, paraffin-embedded human stomach tissue.

  Immunocytochemistry/ Immunofluorescence - HMGB1 antibody (ab77302)

ab77302, at 10µg/ml, staining HMGB1 in Hela cells by Immunofluorescence.

  Flow Cytometry-HMGB1 antibody(ab77302)

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.