Determination of the antibody titer
To determine the titer of the antibody, an ELISA was performed using a serial dilution of antibody directed against DNMT3B (Cat. No. 25219). The plates were coated with the peptides used for immunization of the rabbit. By plotting the absorbance against the antibody dilution (Figure 1), the titer of the antibody was estimated to be 1:220,000.

Western blot analysis using the antibody directed against DNMT3B
Whole cell extracts (25 μg) from HeLa cells were analysed by Western blot using the antibody against DNMT3B (Cat. No. 25219) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.

Immunofluorescence using the antibody directed against DNMT3B
Human HeLa cells were stained with the antibody against DNMT3B (Cat. No. 25219) and with DAPI. Cells were fixed with 4% formaldehyde for 10 minutes and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the DNMT3B antibody (left) diluted 1:1,000 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. 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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