H3K27ac Antibody

Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H3K27ac
ChIP was performed with the Diagenode antibody against H3K27ac (cat. No. C15210016) on sheared chromatin from 500,000 HeLaS3 cells using the “iDeal ChIP-seq” kit (cat. No. C01010051). A titration of the antibody consisting of 0.5, 1, and 2 µg per ChIP experiment was analysed. IgG (1 µg/IP) was used as negative IP control. Quantitative PCR was performed with primers for the promoters of the GAPDH and EIF4A2 genes, used as positive controls, and for the MYOD1 gene and the Sat2 satellite repeat, used as negative controls. The graph shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

Figure 2. Cross reactivity tests using the Diagenode monoclonal antibody directed against H3K27ac
To test the cross reactivity of the Diagenode antibody against H3K27ac (cat. No. C15210016), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H3K27. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:2,000. Figure 2 shows a high specificity of the antibody for the modification of interest.

Figure 3. Western blot analysis using the Diagenode monoclonal antibody directed against H3K27ac
Western blot was performed on whole cell extracts (40 µg) from HeLa cells using the Diagenode antibody against H3K27ac (cat. No. C15210016). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is shown on the right, the marker (in kDa) is shown on the left.

Figure 4. Immunofluorescence using the Diagenode monoclonal antibody directed against H3K27ac
HeLa cells were stained with the Diagenode antibody against H3K27ac (cat. No. C15210016, red) diluted 1:1,000. Actin was stained with fluorescein phalladoin (green).