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Figure 1. ChIP results obtained with the Diagenode antibody directed against FOXM1 ChIP assays were performed using HeLa cells, the Diagenode antibody against FOXM1 (Cat. No. C15410232) 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 CCNA2 and CCNB1 genes, used as positive controls, and for the IFT80 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 FOXM1 ChIP was performed on sheared chromatin from 4 million HeLa cells using 2 μg of the Diagenode antibody against FOXM1 (Cat. No. C15410232) 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 600 kb region of human chromosome 6 (fig 2A and B), and in two genomic regions surrounding the CCNA2 and CCNB1 positive control genes (fig 2C and D).
Figure 3. Western blot analysis using the Diagenode antibody directed against FOXM1 Whole cell extracts from 293T (30 μg, lane 1) or 293T cells transfected with a FOXM1 expression vector (lane 2) were analysed by Western blot using the Diagenode antibody against FOXM1 (Cat. No. C15410232) 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. Immunohistochemistry using the Diagenode antibody directed against FOXM1 Formalin fixed paraffin embedded human ovarian cancer tissue was stained with the Diagenode antibody against FOXM1 (Cat. No. C15410232) diluted 1:100 followed by a peroxidase labelled goat anti-rabbit secondary antibody.
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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