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Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H2A.Z ChIP was performed with the Diagenode antibody against H2A.Z (cat. No. C15210021) 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 2.5 µg per ChIP experiment was analysed. IgG (1 µg/IP) was used as negative IP control. Quantitative PCR was performed with primers for the EIF2S3 and EIF4A2 promoters, used as positive controls, and for the MYOD1 and MYT1 genes, 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. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H2A.Z ChIP was performed on sheared chromatin from 500,000 HeLaS3 cells using 0.5 µg of the Diagenode antibody against H2A.Z (cat. No. C15210021) as described above. The IP'd DNA was subsequently analysed on an Illumina NovaSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the H2A.Z signal along the complete sequence and 1.5 Mb region of the human X-chromosome (figure 2A and B), and in two genomic regions surrounding the EIF2S3 and EIF4A2 positive controls (figure 2C and D).
Figure 3. Western blot analysis using the Diagenode monoclonal antibody directed against H2A.Z Western blot was performed on whole cell extracts (40 µg) from HeLa cells using the Diagenode antibody against H2A.Z (cat. No. C15210021). The antibody was diluted 1:200 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 H2A.Z HeLa cells were stained with the Diagenode antibody against H2A.Z (cat. No. C15210021, red) diluted 1:100. Actin was stained with fluorescein phalladoin (green).
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