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Figure 1. Determination of the antibody titer To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3T45p (cat. No. 15410321) in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 1), the titer of the antibody was estimated to be 1:350,000.
Figure 2. Cross reactivity tests using the Diagenode antibody directed against H3T45p To test the cross reactivity of the Diagenode antibody against H3T45p (cat. No. 15410321), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H3T45. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 2 shows a high specificity of the antibody for the modification of interest.