IHC image of IL6 staining in human lung 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 ab6672, 1/400, 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

Tissue lysates were denatured for 10-15 minutes at 90ºC. ab6672 was incubated overnight at 4ºC and the secondary antibody for 1 hour at RT.

ab6672 detects a band at 25 kDa in Human lung tissue lysate and Mouse spleen tissue lysate, however the signal in mouse tissue is significantly lower. It also binds strongly to a protein at ~55 kDa in Human lung tissue extracts, which we believe represents a glycosylated form of IL6. ab6672 also detects several bands in Human lung tissue lysate within the region of 30-40 kDa. These may represent heteromers of IL6. 

IL6 (Red) and IL-12 (Green) were measured at 1, 3, and 7 days after 10 Gy irradiation of HCa-1 tumors to determine whether irradiation regulates IL-12 and IL6 expression in tumours. ab6672 was used to stain IL6 at 1/100 dilution in immunohistochemical analysis.

4-20% Tris-Glycine gel.

The membrane was blocked for 30 minutes with 1% BSA-TBST.