Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Rat cerebrum tissue labelling BAP1 with ab255611 at 1/100 dilution. Heat mediated antigen retrieval was performed usingTris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins, LeicaDS9800 (Bond™ Polymer Refine Detection) was used as the secondary antibody.  Negative control using PBS instead of primary antibody. Counterstained with hematoxylin. Nuclear staining on rat cerebrum. The section was incubated with ab255611 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse cerebrum tissue labelling BAP1 with ab255611 at 1/100 dilution. Heat mediated antigen retrieval was performed usingTris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins, LeicaDS9800 (Bond™ Polymer Refine Detection) was used as the secondary antibody.  Negative control using PBS instead of primary antibody. Counterstained with hematoxylin. Nuclear staining on mouse cerebrum. The section was incubated with ab255611 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human lung carcinoma tissue labelling BAP1 with ab255611 at 1/100 dilution. Heat mediated antigen retrieval was performed usingTris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins, LeicaDS9800 (Bond™ Polymer Refine Detection) was used as the secondary antibody.  Negative control using PBS instead of primary antibody. Counterstained with hematoxylin. Nuclear staining on human lung carcinoma. The section was incubated with ab255611 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human colon carcinoma tissue labelling BAP1 with ab255611 at 1/100 dilution. Heat mediated antigen retrieval was performed usingTris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins, LeicaDS9800 (Bond™ Polymer Refine Detection) was used as the secondary antibody.  Negative control using PBS instead of primary antibody. Counterstained with hematoxylin. Nuclear staining on human colon carcinoma. The section was incubated with ab255611 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

All lanes : Anti-BAP1 antibody [EPR22826-65] (ab255611) at 1/1000 dilution

Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate
Lane 2 : PC-3 (human prostate adenocarcinoma epithelial cell), whole cell lysate
Lane 3 : HepG2 (human hepatocellular carcinoma epithelial cell), whole cell lysate
Lane 4 : A375 (human malignant melanoma epithelial cell), whole cell lysate
Lane 5 : MDA-MB-231 (human breast adenocarcinoma epithelial cell), whole cell lysate
Lane 6 : A549 (human lung carcinoma epithelial cell), whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 80 kDa
Observed band size: 100 kDa why is the actual band size different from the predicted?


Exposure time: 3 minutes

The molecular weight observed is consistent with what has been described in the literature (PMID: 18757409).

Blocking/Dilution buffer: 5% NFDM/TBST.

Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized PC-3 (human prostate adenocarcinoma epithelial cell) cells labeling BAP1 with ab255611 at 1/60 (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.