RNF2 Antibody - ChIP-seq Grade

Figure 1. ChIP results obtained with the Diagenode antibody directed against RNF2
ChIP assays were performed using K562 cells, the Diagenode antibody against RNF2 (cat. No. C15410313) and optimized PCR 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 consisting of 1, 2 and 5 μg of antibody per ChIP experiment was analyzed. IgG (2 μg/IP) was used as a negative IP control. Quantitative PCR was performed with optimized primers for the ARNTL2 gene and the non coding RNA genes LINC00629 and LINC00875, used as positive controls, and for the EIF4A2 promoter, used as a negative control. 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 RNF2
ChIP was performed on sheared chromatin from 4,000,000 K562 cells using 2 μg of the Diagenode antibody against RNF2 (Cat. No. C15410313) as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq 2000. 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 peak distribution along the complete sequence and a 300 kb region surrounding the HOX cluster of chromosome 7 (figure 2A and B) and in two regions surrounding the ARNTL2 and LINC00629 positive control genes, respectively (figure 2C and D). The position of the amplicon used for ChIP-qPCR is indicated by an arrow.

Figure 3. Western blot analysis using the Diagenode antibody directed against RNF2
Whole cell extracts from Jurkat cells (lane 1) and mouse testis (lane 2) were analysed by Western blot using the Diagenode antibody against RNF2 (Cat. No. C15410313) diluted 1:1,000. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.

Figure 4. Immunofluorescence using the Diagenode antibody directed against RNF2
U2OS cells were stained with the Diagenode antibody against RNF2 (Cat. C15410313) diluted 1:200 (left). The right panel shows costaining of the nuclei with DAPI.