All lanes : HRP Anti-Bcl-2 antibody [EPR17509] (ab209039) at 1/5000 dilution

Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : BCL2 knockout HAP1 whole cell lysate

Lysates/proteins at 20 µg per lane.

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 26 kDa


Exposure time: 20 minutes

ab209039 was shown to specifically react with Bcl-2 in wild-type HAP1 cells as signal was lost in BCL2 knockout cells. Wile-type and BCL2 knockout samples were subjected to SDS-PAGE. Ab209039 was incubated overnight at 4°C at 1/5000 dilution. Blots were developed with ECL technique.

IHC image of Bcl-2 staining in a section of formalin-fixed paraffin-embedded normal human spleen*, performed on a Leica BOND™. The section was pre-treated using heat mediated antigen retrieval with Tris/EDTA buffer (pH9, epitope retrieval solution 2) for 20mins. The section was then incubated with ab209039, 1/250 dilution, for 15 mins at room temperature. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset negative control image is taken from an identical assay without primary antibody.

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.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

All lanes : HRP Anti-Bcl-2 antibody [EPR17509] (ab209039) at 1/5000 dilution

Lane 1 : THP1 (Human acute monocytic leukemia cell line) Whole Cell Lysate
Lane 2 : Tonsil (Human) Tissue Lysate - adult normal tissue
Lane 3 : Thymus (Human) Tissue Lysate - adult normal tissue
Lane 4 : Spleen (Mouse) Tissue Lysate

Lysates/proteins at 10 µg per lane.

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 26 kDa
Observed band size: 26 kDa


Exposure time: 8 minutes

This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab209039 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.