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Our Abpromise guarantee covers the use of ab5176 in the following tested applications.
|ICC/IF||Use at an assay dependent concentration.|
|WB||1/500 - 1/1000. Detects a band of approximately 17 kDa (predicted molecular weight: 17 kDa).Can be blocked with Human Histone H3 (phospho S10) peptide (ab11477).|
|IHC-P||Use a concentration of 5 µg/ml.|
|IHC - Wholemount||1/250. PubMed: 20144789|
|IHC-Fr||1/200. (see Abreview)|
ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.
|ChIP||Use at an assay dependent concentration. PubMed: 20089855|
|IHC (PFA fixed)||Use at an assay dependent concentration. PubMed: 25359725|
|IHC-FoFr||Use at an assay dependent concentration. PubMed: 19622634|
IHC image of ab5176 staining Histone H3 (phospho S10) in human kidney 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 ab5176, 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.
Lysates: Lane 1,3,5: 0.5µg/lane untreated histone Lane 2,4,6: 0.5µg/lane colcemid treated histone - treated Primary antibody: Lane 1-2 : ab5176 (1?g/ml) Lane 3-4 :ab5176 (1?g/ml) + phospho peptide (ab5176) 1?g/ml Lane 5-6 :ab5176 (1?g/ml) + non-phospho peptide ( ab2903 batch 3) 1?g/ml