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Synthetic peptide conjugated to KLH derived from within amino acid residues 100 to the C-terminus of Human H2A.X.
Our Abpromise guarantee covers the use of ab20669 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa).Can be blocked with Human gamma H2A.X (unmodified ) peptide (ab15646).|
|IP||Use at an assay dependent concentration.|
|ICC/IF||Use at an assay dependent concentration.|
|IHC-P||Use a concentration of 0.2 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
ab20669 staining Histone H2A.X in Mouse brain neurone cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.3% Triton X-100 and blocked with 5% serum for 2 hours at 22°C. Samples were incubated with primary antibody (1/500 in 2.5% serum + 0.3% Triton X-100 in PBS) for 24 hours at 4°C. ab175651, an Alexa Fluor® 405-conjugated Donkey anti-rabbit IgG polyclonal (1/500) was used as the secondary antibody.
IHC image of Histone H2A.X 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 ab20669, 0.2 µg/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.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"