Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse cerebrum tissue sections labeling HDAC3 with Purified ab76295 at 1:100 dilution (1.03 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody. Negative control:PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling HDAC3 with Purified ab76295 at 1:20 dilution (10 µg/ml) (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1:2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling HDAC3 with Purified ab76295 at 1:50 dilution (2.1 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Rat kidney tissue sections labeling HDAC3 with Purified ab76295 at 1:100 dilution (1.03 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody. Negative control:PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human lung carcinoma tissue sections labeling HDAC3 with Purified ab76295 at 1:100 dilution (1.03 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody. Negative control:PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

All lanes : Anti-HDAC3 antibody [EP462Y] (ab76295) at 1/2000 dilution

Lane 1 : HeLa cell lysate
Lane 2 : Jurkat cell lysate
Lane 3 : HepG2 cell lysate
Lane 4 : NIH/3T3 cell lysate
Lane 5 : PC-12 cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 49 kDa
Observed band size: 49 kDa

Anti-HDAC3 antibody [EP462Y] (ab76295) at 1/2000 dilution + HeLa cell lysate at 10 µg

Secondary
goat anti-rabbit HRP at 1/2000 dilution

Predicted band size: 49 kDa
Observed band size: 49 kDa

ab76295, at a 1/100-1/250 dilution, staining human HDAC3 in breast carcinoma, by Immunohistochemistry, Formalin/PFA-fixed paraffin-embedded tissue.

Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.