Recombinant HRP Anti-MEK1 antibody [E342] (ab193987)

ab193987 was shown to recognize MEK1 in wild-type HAP1 cells as signal was lost at the expected MW in MAP2K1 (MEK1) knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and MAP2K1 (MEK1) knockout samples were subjected to SDS-PAGE. Ab193987 and ab184095 (Mouse monoclonal [mAbcam 9484] to GAPDH - Loading Control (Alexa Fluor^® 680) loading control) were incubated overnight at 4°C at 1/5000 dilution and 1/1000 dilution respectively. The loading control was imaged using the Licor Odyssey CLx prior to blots being developed with ECL technique.

IHC image of MEK1 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 ab193987 at a working dilution of 1/500. 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

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 2% Bovine Serum Albumin before being incubated with ab193987 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.