HRP Anti-Transferrin antibody [EPR2932(2)] - Serum Loading Control (ab185070) at 1/5000 dilution + Human prostate tumor tissue lysate (adenocarcinoma) - total protein (ab30305) at 10 µg

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 77 kDa
Observed band size: 82 kDa why is the actual band size different from the predicted?


Exposure time: 30 seconds

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 5% Bovine Serum Albumin before being incubated with ab185070 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.

IHC image of Transferrin staining in a section of formalin-fixed paraffin-embedded cancerous human liver*. 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 ab185070 at 10µg/ml. 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