ChIP results obtained with the monoclonal antibody directed against HDAC3
ChIP assays were performed using human HeLa cells, the monoclonal antibody against HDAC3 (Cat. # 25289) and optimized PCR primer sets for qPCR. ChIP was performed on sheared chromatin from 10,000 cells. A titration of the antibody consisting of 1, 2, 5, and 10 µg per ChIP experiment was analyzed. IgG (5 µg/IP) was used as negative IP control. QPCR was performed with primers for the promoters of the active genes c-fos and GAPDH, and for the coding region of p21, a known target gene of HDAC3. Figure 4 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

Immunofluorescence using the monoclonal antibody directed against HDAC3
HeLa cells were stained with the antibody against HDAC3 (Cat. # 25289) and with DAPI. Cells were fixed with 4% formaldehyde for 10 min. and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the HDAC3 antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.

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