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Our Abpromise guarantee covers the use of ab7028 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/2000 - 1/20000. Predicted molecular weight: 55 kDa.|
|IP||Use at an assay dependent concentration.|
|IHC-P||1/500. Perform enzymatic antigen retrieval before commencing with IHC staining protocol.|
|ChIP/Chip||Use at an assay dependent concentration.|
|ChIP||Use 5µl for 106 cells.|
|ICC/IF||Use at an assay dependent concentration.|
|IHC-Fr||Use at an assay dependent concentration. PubMed: 23911933|
Sonicated Chromatin prepared from untreated (UI) or 17beta-estradiol (E2) treated MCF7 cells was subjected to the ChIP procedure with ab7028 to HDAC1 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). 5 µl of ab7028 and 2x10.
Immunofluorescence analysis of methanol fixed HeLa cells with ab7028 labelling HDAC1 at 1:200 dilution. The antibody was developed using Goat Anti-Rabbit IgG, Cy3 conjugate.
Immunohistochemical analysis of paraformaldehyde fixed paraffin embedded human breast tumour section with ab7028 labelling HDAC1 at 1/500 dilution.
HDAC1 was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to HDAC1 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab7028.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 60ka: HDAC1.
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