Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human endometrial adenocarcinoma tissue labelling gamma H2A.X with ab195190 at 5 µg/mL. Heat mediated antigen retrieval was performed using Bond™ Epitope Retrieval Solution 1 (pH 6.0) for 20 minutes. Negative control using PBS instead of primary antibody. Counterstained with hematoxylin. 

Nuclear staining on human endometrial adenocarcinoma.

The section was incubated with ab195190 for 15 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument

 

All lanes : HRP Anti-gamma H2A.X (phospho S139) antibody [EP854(2)Y] (ab195190) at 1/10000 dilution

Lane 1 : Untreated Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate
Lane 2 : Jurkat (Human T cell leukemia T lymphocyte) treated with 25µM Etoposide for 8 hours whole cell lysate

Lysates/proteins at 15 µg per lane.

Predicted band size: 15 kDa
Observed band size: 15 kDa


Exposure time: 15 seconds

Blocking/diluting buffer and concentration: 5% NFDM/TBST

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human testis tissue labelling gamma H2A.X with ab195190 at 5 µg/mL. Heat mediated antigen retrieval was performed using Bond™ Epitope Retrieval Solution 1 (pH 6.0) for 20 minutes. Negative control using PBS instead of primary antibody. Counterstained with hematoxylin. 

Nuclear staining on human testis.

The section was incubated with ab195190 for 15 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument

IHC image of Histone H2A.X staining in a section of formalin-fixed paraffin-embedded normal human colon*, 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 ab195190, 1/71.4285714285714 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