HDAC1 monoclonal antibody

Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against HDAC1
ChIP assays were performed using human HeLa cells, the Diagenode monoclonal antibody against HDAC1 (Cat. No. C15200144) 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 GAPDH promoter and for the coding region of p21, a known target gene of HDAC1. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

Figure 2. Western blot analysis using the Diagenode monoclonal antibody directed against HDAC1
Nuclear extracts from HeLa cells (40 μg) were analysed by Western blot using the Diagenode monoclonal antibody against HDAC1 (Cat. No. C15200144) diluted 1:2,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right (expected size: 55 kDa); the marker (in kDa) is shown on the left.

Figure 3. Western blot analysis using the Diagenode monoclonal antibody directed against HDAC1
Whole cell extracts (40 μg) from HeLa cells transfected with HDAC1 siRNA (lane 2) and from an untransfected control (lane 1) were analysed by Western blot using the Diagenode antibody against HDAC1 (Cat. No. C15200144) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right (expected size: 55 kDa); the marker (in kDa) is shown on the left.

Figure 4. Immunofluorescence using the Diagenode monoclonal antibody directed against HDAC1
HeLa cells were stained with the Diagenode antibody against HDAC1 (Cat. No. C15200144) 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 HDAC1 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.