IHC image of PSMB10 staining in Human colon adenocarcinoma formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab77735, 10µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
Anti-PSMB10 antibody (ab77735) at 5 µg/ml + Recombinant tagged human PSMB10 protein at 0.2 µgSecondaryGoat Anti-Mouse IgG (H&L)-HRP Conjugate at 1/5000 dilution
Anti-PSMB10 antibody (ab77735) at 5 µg/ml + Jurkat cell lysate at 25 µgSecondaryGoat Anti-Mouse IgG (H&L)-HRP Conjugate at 1/2500 dilution
ab77735, at 10µg/ml, staining PSMB10 in Hela cells by Immunofluorescence.
Overlay histogram showing HeLa cells stained with ab77735 (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 (ab77735, 1µg/1x106 cells) 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.