IHC image of PGK1 staining in a section of formalin-fixed paraffin-embedded human normal colon*. The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH6) for 30mins, and incubated overnight at +4°C with ab197960 at 1/71 dilution. DAB was used as the chromogen (ab103723), diluted 1/100 and incubated for 10min at room temperature. The section was 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

HRP Anti-PGK1 antibody [22C5D8] (ab197960) at 1/5000 dilution + S.cerevisiae (Y190) Whole Cell Lysate at 20 µg

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 45 kDa
Observed band size: 45 kDa


Exposure time: 1 minute

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 ab197960 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.