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Figure 1. ChIP results obtained with the Diagenode antibody directed against NF-E2 ChIP assays were performed using K562 cells, the Diagenode antibody against NF-E2 (Cat. No. C15410240) 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 HBE1 and GAPDH 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 NF-E2 ChIP was performed on sheared chromatin from 5 million K562 cells using 5 μg of the Diagenode antibody against NF-E2 (Cat. No. C15410240). 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 Mb region of the human X chromosome (fig 2A and B), and in a two genomic regions of chromosome 11 and 12 surrounding the HB cluster and the GAPDH genes, respectively.
Figure 3. Western blot analysis using the Diagenode antibody directed against NF-E2 Whole cell extracts from 293 cells (30 μg, figure 3A) or mouse spleen (50 μg, figure 3B) were analysed by Western blot using the Diagenode antibody against NF-E2 (Cat. No. C15410240) 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 with the Diagenode antibody directed against NF-E2 HepG2 cells were fixed with 4% formaldehyde for 15’ at room temperature and stained with the Diagenode antibody against NF-E2 (Cat. C15410240) diluted 1:500 (left). The right picture shows costaining with Hoechst 33342 nucleic acid stain.
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Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9
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WB Western blot : The quality of antibodies used in this technique is crucial for correct and specific protein identification. Diagenode offers huge selection of highly sensitive and specific western blot-validated antibodies.
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