H3K36me2 Antibody - ChIP Grade (sample size)

Figure 1. ChIP results obtained with the monoclonal antibody directed against H3K36me2
ChIP assays were performed using human HeLa cells, a monoclonal antibody against H3K36me2 (cat. No. C15200182), and optimized PCR primer pairs for qPCR. ChIP was performed on sheared chromatin from 1 million cells. A titration consisting of 0.5, 1, 2 and 5 µg of antibody per ChIP experiment was analyzed. IgG (1 µg/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for a genomic region upstream of the TGM2 gene, used as a positive control, and for the inactive MYOD1 gene and the Sat2 satellite repeat, used as negative controls. 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. Cut&Tag results obtained with the monoclonal antibody directed against H3K36me2
Cut&Tag was performed on 50,000 K562 cells using 1 µg of the monoclonal antibody against H3K36me2 (cat. No. C15200182), the pA-Tn5 transposase (C01070001) and the iDeal Cut&Tag kit (cat. No. C01070021). The libraries were subsequently analyzed on an Illumina NovaSeq sequencer (2x50 paired-end reads) according to the manufacturer’s instructions. The tags were aligned to the human genome (hg38) using the BWA algorithm. Figure 2 shows the peak distribution in two genomic regions on chromosome 3 and X (figure 2A and 2B, respectively).

Figure 3. Cross reactivity of the monoclonal antibody directed against H3K36me2
To test the specificity, an ELISA was performed using a serial dilution of the monoclonal antibody against H3K36me2 (cat. No. C15200182). The wells were coated with peptides containing the unmodified H3K36 region as well as the mono-, di-, and trimethylated H3K36 and the trimethylated H3K9. Figure 3 shows a high specificity of the antibody for the peptide containing the modification of interest.