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Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against HDAC3 ChIP assays were performed using human HeLa cells, the Diagenode monoclonal antibody against HDAC3 (Cat. No. C15200145) and optimized PCR primer sets for qPCR. ChIP was performed with the “LowCell# ChIP” kit (cat. No. kch-maglow-016), on sheared chromatin from 10,000 cells using the SX-8G IP-Star automated system. A titration of the antibody consisting of 1, 2, 5, and 10 μg per ChIP experiment was analysed. 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).
Figure 2. Immunofluorescence using the Diagenode monoclonal antibody directed against HDAC3 HeLa cells were stained with the Diagenode antibody against HDAC3 (Cat. No. C15200145) and with DAPI. Cells were fixed with 4% formaldehyde for 10’ 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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Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9
HeLa cells transfected with a Cas9 expression vector (... Read more