IHC image of ab100944 staining Histone H1.01 in chicken colon formalin fixed paraffin embedded tissue sections, performed on a Leica Bond. 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 ab100944, 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. No primary antibody was used in the negative control (shown on the inset).
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.
Western blot - Histone H1.01 antibody - ChIP Grade (ab100944)
Competition Western Blot Analysis. The recombinant linker histone was separated on a 15% SDS PAGE slab gel together with a track loaded with total chicken histone as marker and transferred to nitrocellulose membranes. These were then cut into strips of equal width and each strip developed with the antibody which had been pre-incubated with one of the recombinant linker histone at four increasing concentrations (two are shown). The panels show (from left to right): the transfer of the recombinant protein slab gel; the transfer of a total chicken histone marker, M; the histone marker strip blotted with the antibody under test, M(WB), showing that there is no recognition of the core histones, nor H5, with the antibody; six strips developed with the test antibody (at 0.1 mg/ml) pre-incubated with the six recombinant histones (01 to R) at the indicated concentrations. The Control lanes represent blots of a seventh strip developed with the test antibody without any competitor. The red boxes