HRP Anti-pro Caspase-3 antibody [E61]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - HRP Anti-pro Caspase-3 antibody [E61] (AB205733)

IHC image of pro Caspase-3 staining in a section of formalin-fixed paraffin-embedded normal human tonsil*, performed on a Leica BOND™. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab205733, 1/2500 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

• WB

Lab

Western blot - HRP Anti-pro Caspase-3 antibody [E61] (AB205733)

This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 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 ab205733 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.

All lanes:

Western blot - HRP Anti-pro Caspase-3 antibody [E61] (ab205733) at 1/5000 dilution

Lane 1:

Western blot - HeLa whole cell lysate (<a href='/en-us/products/cell-lysates/hela-whole-cell-lysate-ab150035'>ab150035</a>) at 10 µg

Lane 2:

Wild-type HAP1 cell lysate at 20 µg

Lane 3:

Wild-type HAP1 cell lysate Stuarosporine Treated (1μM for 4h) at 20 µg

Lane 4:

Caspase-3 knockout HAP1 cell lysate at 20 µg

Lane 5:

Caspase-3 knockout HAP1 cell lysate Stuarosporine Treated (1μM for 4h) at 20 µg

Predicted band size: 31 kDa

Observed band size: 35 kDa

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Exposure time: 3min