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Figure 1. ChIP results obtained with the Diagenode antibody directed against SMAD1(S206p) ChIP assays were performed using K562 cells, the Diagenode antibody against SMAD1(S206p) (Cat. No. C15410274) 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 EIF4A2 and GAPDH promoters, 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 SMAD1(S206p) ChIP was performed on sheared chromatin from 5 million K562 cells using 5 μg of the Diagenode antibody against SMAD1(S206p) (Cat. No. C15410274). 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 3 Mb region of the human X chromosome (fig 2A and B) and in two genomic regions surrounding the GAPDH and EIF4A2 genes on chromosome 12 and 3, respectively (fig 2C and D).
Figure 3. Western blot analysis using the Diagenode antibody directed against SMAD1(S206p) Whole cell extracts (15 μg) from HepG2 (lane 1) and HaCat (lane 2) cells were analysed by Western blot using the Diagenode antibody against SMAD1(S206p) (Cat. No. C15410274) diluted 1:500.
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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