Figure 1. hMeDIP with the Diagenode rat IgG negative control antibody
hMeDIP assays were performed using the Diagenode rat monoclonal antibody against 5-hmC (cat. No. C15220001) and the “hMeDIP” kit (rat, cat. No. C02010030) on 1 μg of sheared DNA from mouse ES cells. The DNA was spiked with 0.025 ng of each of the DNA standards from the 5-hmC, 5-mC & cytosine DNA standard pack (cat. No. C02040011), prior to the IP Rat IgG (cat. No. C15420001) was used as a negative IP control. 2.5 μg of antibody per ChIP experiment was used for both antibodies. Quantitative PCR was performed with the primers specific for the unmethylated, methylated and hydroxymethylated controls present in the kit. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).