IHC image of CHD4 staining in human breast carcinoma formalin fixed paraffin embedded tissue section*, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab70469, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
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
Immunocytochemical analysis of blastocysts from Wildtype and CHD4 -/- (Knockout) mice, labeling CHD4 with ab70469. Embryos were fixed in 2.5% paraformaldehyde, permeablised with 0.25% Triton X-100 and 3% polyvinlypyrrolidone in PBS, then blocked in 10% foetal calf serum and 0.1 Triton X-100 in PBS. Immunostaining with ab70469 (diluted 1/10,000) was performed overnight at 4°C. Staining with secondary antibody was for 1 hour at RT. DAPI used for nuclear staining (blue).
Western blot - Anti-CHD4 antibody [3F2/4] - ChIP Grade (ab70469)
All lanes : Anti-CHD4 antibody [3F2/4] - ChIP Grade (ab70469) at 5 µg/ml
Lane 1 : Human kidney tissue lysate - total protein (ab30203) Lane 2 : Kidney (Mouse) Tissue Lysate Lane 3 : Human placenta tissue lysate - total protein (ab29745) Lane 4 : Human skin tissue lysate - total protein (ab30166) Lane 5 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 218 kDa Additional bands at: 238 kDa. We are unsure as to the identity of these extra bands.