All lanes : Anti-IRE1 antibody [9F2] (ab96481) at 1/500 dilutionLane 1 : Raji cell lysateLane 2 : A431 cell lysateLane 3 : Jurkat cell lysateLane 4 : HeLa cell lysateLane 5 : HEK293 cell lysate
Immunohistochemical analysis of paraffin-embedded Human brain tissue (A) and stomach tissue (B), showing cytoplasmic localization using ab96481 at 1/200 dilution with DAB staining.
Overlay histogram showing Raji cells stained with ab96481 (red line). The cells were fixed with 80% 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 (ab96481, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was 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 Raji cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.