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ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.
Our Abpromise guarantee covers the use of ab1764 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ChIP||Use at an assay dependent concentration.|
|ELISA||Use at an assay dependent concentration.|
|IP||Use at an assay dependent concentration.|
|WB||1/800 - 1/2000. Detects a band of approximately 20 kDa.
Use 5% BSA in TBST to block.
|IHC-P||1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
|ICC/IF||Use at an assay dependent concentration.|
IHC image of ab1764 staining Histone H2A (acetyl K5) in human 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 ab1764, 1/1000 dilution, 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.
IHC - Wholemount of Caenorhabditis elegans whole body labelling Histone H2A (acetyl K5) with ab1764. Sample was incubated with primary antibody (1/2500 in 3% BSA + 0.1% Triton X-100 in PBS) for 12 hours at 4°C. An Alexa Fluor® 488-conjugated Goat anti-rabbit IgG polyclonal (1/1000) was used as the secondary antibody. Left - Histone H2A (acetyl K5) (green), Right - nuclei (DAPI, blue).
ab1764 staining Female Embryonic Stem cells differentiated for 7 days by immunocytochemistry. The antibody was used at a dilution of 1/200 and incubated with the cells for 1 hour. Bound antibody was detected with a FITC conjugated Goat anti-Rabbit polyclonal antibody.
DAPI counterstain is shown pseudocoloured as white on the right of the image (b). Centric heterochromatin was underacetylated in differentiated cells (arrowhead in a). A pale staining chromosome was observed at high frequencies; previoulsy shown to be the inactive female X chromosome (arrow in a).
This image is courtesy of an Abreview by Hugh Spotswood submitted on 13 February 2006.
This image is courtesy of an anonymous AbreviewAb1764 at a 1/1000 dilution in Western blot of human Osteosarcoma (U2OS) nuclear cell lysate.
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