TATA binding protein TBP Antibody (C.15200002)

Western Blot: TATA binding protein TBP Antibody (C.15200002) [NBP2-59208]

Western Blot: TATA binding protein TBP Antibody (C.15200002) [NBP2-59208] - Whole cell extracts (40 ug) from HeLa cells transfected with TBP siRNA (lane 2) and from an untransfected control (lane 1) were analysed by Western blot using the antibody against TBP diluted 1:500 in TBSTween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (kDa) is shown on the left.

Chromatin Immunoprecipitation Sequencing: TATA binding protein TBP Antibody (C.15200002) [NBP2-59208] - IP'd DNA was analyzed by QPCR with optimized PCR primer pairs for the promoters of the active GAPDH and c-fos genes, used as positive control targets, and for a region 1 kb upstream of the GAPDH promoter and the coding region of the inactive MB gene, used as negative control targets (A). The IP'd DNA was analyzed with an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. Figure shows the peak distribution in 50 kb regions surrounding the GAPDH, c-fos, ACTB and MCL1 genes (B, C, D and E, respectively). These results clearly show a localisation of TBP at the promoters of actively transcribed genes.

Chromatin Immunoprecipitation: TATA binding protein TBP Antibody (C.15200002) [NBP2-59208] - ChIP results obtained with the antibody against TBP. ChIP assays were performed using U2OS cells, the antibody were used per ChIP experiment. QPCR was performed with primers for the promoter of the c-fos and GAPDH genes, a region 0.5 and 1 kb upstream of the GAPDH promoter, respectively, and for exon 2 of the myoglobin gene as a negative control. Figure shows the recovery (the relative amount of immunoprecipitated DNA compared to input DNA) and the occupancy (ratio +/- control target). These results demonstrate the occupancy of both promoters by TBP.