GTF2E2 polyclonal antibody - Classic

Figure 1. ChIP results obtained with the Diagenode antibody directed against GTF2E2
ChIP assays were performed using HeLa cells, the Diagenode antibody against GTF2E2 (Cat. No. C15410264) and optimized primer sets for qPCR. ChIP was performed with the “iDeal ChIP- seq” kit (Cat. No. C01010055), using sheared chromatin from 4 million cells. A titration of the antibody consisting of 1, 2 and 5 μg per ChIP experiment was analysed. IgG (1 μg/IP) was used as negative IP control. QPCR 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. 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. ChIP-seq results obtained with the Diagenode antibody directed against GTF2E2
ChIP was performed on sheared chromatin from 4 million HeLa cells using 5 μg of the Diagenode antibody against GTF2E2 (Cat. No. C15410264) as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the ChIP-seq signal in three genomic regions surrounding the GAPDH and EIF4A2 positive control genes (fig 2A and B) and the EIF3S2 gene (fig 2C).

Figure 3. Western blot analysis using the Diagenode antibody directed against GTF2E2
Whole cell extracts from Jurkat cells (30 μg) were analysed by Western blot using the Diagenode antibody against GTF2E2 (Cat. No. C15410264) diluted 1:5,000. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.

Figure 4. Immunoprecipitation using the Diagenode antibody directed against GTF2E2
Immunoprecipitation was performed on whole cell extracts from Jurkat cells using 2.5 μg of the Diagenode antibody against GTF2E2 (Cat. No. C15410264). An equal amount of rabbit IgG was used as a negative control. The immunoprecipitated GTF2E2 protein was detected by western blot with the GTF2E2 antibody diluted 1:1,000. The IP with the GTF2E2 antibody and with the IgG negative control are shown in lane 3 and lane 2, respectively. Lane 1 shows the input (30 μg of Jurkat whole cell extract).