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Synthetic peptide conjugated to KLH derived from within residues 50 - 150 of Human BAF53A.
(Peptide available as ab13770.)
Our Abpromise guarantee covers the use of ab3882 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.|
|ICC/IF||Use at an assay dependent concentration.|
|WB||1/1000. Detects a band of approximately 53 kDa.|
|IHC-P||1/35. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
Total extracts from two different clones of MCF7 cells expressing a TAP tagged version of Tip60 were partially purified over IgG sepharose resin and eluted with TEV protease. Western blot was performed using a 1/1000 dilution of the BAF53A antibody.
Lane 1. TEV elution of Tip60-TAP clone #1
Lane 2. TEV elution of Tip60-TAP clone #2
Lane 3. Total extract of Tip60-TAP clone #1
Lane 4. Total extract of Tip60-TAP clone #2
NB: Tip60 is detected in the total extract due to the presence of Protein A in the TAP Tag (to which the primary and secondary abs bind). The TEV protease removes the Tag in lanes 1 and 2.
Sonicated Chromatin prepared from untreated (UI) or 17beta-estradiol (E2) treated MCF7 cells was subjected to the ChIP procedure with ab3882 to BAF53A and the immunoprecipitated chromatin was analysed in the proximal region of the estrogen-responsive pS2 promoter (as shown above) and quantified by real-time PCR (values are nomalized over inputs). The primers are designed to follow the nucleosome E (including the Estrogen Responsive Element ERE). 8 µl of ab3882 and 2x106 cells were used in each ChIP experiment.
ab3882 at a 1/200 dilution staining asynchronous HeLa cells by ICC/IF. The cells were paraformaldehyde fixed and incubated with the antibody for 30 minutes at room temperature. Bound antibody was detected using a Cy3 conjugated Goat anti-rabbit antibody. Nuclei were visualised using DAPI stain. ab3882 gives a predominantly nuclear staining pattern.
This image is courtesy of an Abreview submitted by Kirk McManus.
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