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C-KEEKPEAKGVKEEVKLAconjugated to KLH, corresponding to amino acids 466-482 of Human HDAC 1.
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.
|Flow Cyt||Use at an assay dependent concentration.|
|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|
HeLa cells were fixed and permeabilized with cold methanol followed by cold methanol:acetone. Fixed cells were stained with 1:200 ab7028. The antibody was developed using Goat Anti-Rabbit IgG, Cy3 conjugate.
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.
This image is courtesy of an anonymous Abreview
This image is courtesy of an anonymous AbreviewBlocked with 5% milk for 1 hour at 20°C
ab7028 staining HDAC1 in human primary cardiac cells by Flow Cytometry. Cells were fixed with paraformaldehyde and permeabilized with permeabilization buffer. The sample was incubated with the undiluted primary antibody for 12 hours at 4°C. An Allophycocyanin-conjugated goat polyclonal anti-rabbit IgG (1/1000) was used as the secondary antibody.
Gating Strategy: Isotype negative control (white).
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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