ChIP results obtained with the monoclonal antibody directed against HDAC1
ChIP assays were performed using human HeLa cells, the monoclonal antibody against HDAC1 (Cat. # 25286) 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 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).

Western blot analysis using the monoclonal antibody directed against HDAC1
Nuclear extracts from HeLa cells (40 µg) were analyzed by Western blot using the monoclonal antibody against HDAC1 (Cat. # 25286) 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.

Immunofluorescence using the monoclonal antibody directed against HDAC1
HeLa cells were stained with the antibody against HDAC1 (Cat. # 25286) 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 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.

Store at –80°C for up to 2 years. Centrifuge after first thaw to maximize product recovery. Aliquot to avoid repeated freeze/thaw cycles. Aliquots may be stored at –20°C for at least one month.