SIN3A polyclonal antibody - Classic

Figure 1. ChIP results obtained with the Diagenode antibody directed against SIN3A
ChIP assays were performed using HeLa cells, the Diagenode antibody against SIN3A (Cat. No. C15410250) 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 EIF2S3 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 SIN3A
ChIP was performed on sheared chromatin from 4 million HeLa cells using 2 μg of the Diagenode antibody against SIN3A (Cat. No. C15410250) 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 enrichment along the complete sequence and a 1.5 Mb region of human chromosome 1 (fig 2A and B), and in two genomic regions surrounding the EIF2S3 and GAPDH positive control genes (fig 2C and D).

Figure 3. Western blot analysis using the Diagenode antibody directed against SIN3A
Whole cell extracts from 293T (lane 1), A431 (lane 2), HeLa (lane 3) and HepG2 (lane 4) cells were analysed by Western blot using the Diagenode antibody against SIN3A (Cat. No. C15410250) 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. Immunoprecipitation using the Diagenode antibody directed against SIN3A
Immunoprecipitation was performed on whole cell extracts from 293T cells using 3 μg of the Diagenode antibody against SIN3A (Cat. No. C15410250). An equal amount of rabbit IgG was used as a negative control. The immunoprecipitated SIN3A protein was detected by western blot with the SIN3A antibody diluted 1:1,000. The IP with the SIN3A antibody and with the IgG negative control are shown in lane 3 and lane 2, respectively. Lane 1 shows the input (30 μg of 293T whole cell extract).

Figure 5. Immunofluorescence with the Diagenode antibody directed against SIN3A
HeLa cells were fixed with formaldehyde and stained with the Diagenode antibody against SIN3A (Cat. C15410250) diluted 1:1,000 (left). The right picture shows costaining with Hoechst 33342 nucleic acid stain.

Figure 6. Immunohistochemistry using the Diagenode antibody directed against SIN3A
Formalin fixed paraffin embedded mouse prostate (fig 6A), mouse colon (fig 6B) or rat fore brain (fig 6C) tissue was stained with the Diagenode antibody against SIN3A (Cat. No. C15410250) diluted 1:500 followed by a peroxidase labelled goat anti-rabbit secondary antibody.