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Applications: ChIP/ChIP - seq (4 - 5 µg/IP)
WB (1:200 - 1:2000)
Assay Conditions: ChIP results obtained with the monoclonal antibody against TBP
ChIP assays were performed using U2OS cells, the antibody directed against TBP (Cat. # 25319) and optimized primer sets for qPCR. Sheared chromatin from 1x10^6 cells and 4 µg of 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 1 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.ChIP-seq results obtained with the monoclonal antibody directed against TBP
ChIP was performed with 5 µg of the antibody against TBP (Cat. # 25319) on sheared chromatin from 1 million HeLaS3 cells. The immunoprecipitated 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 (figure 2A). The immunoprecipitated DNA was subsequently 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 2 shows the peak distribution in 50 kb regions surrounding the GAPDH, c-fos, ACTB and MCL1 genes (figure 2B, C, D and E, respectively). These results clearly show a localisation of TBP at the promoters of actively transcribed genes.
Background: TATA-binding protein (TBP) is a transcription factor that binds specifically to a DNA sequence called the TATA box. This DNA sequence is found approximately 30 base pairs upstream of the transcription start site in 10-20% of eukaryotic gene promoters. TBP is a subunit of the eukaryotic transcription factor TFIID. TBP is also a component of RNA polymerase I and RNA polymerase III, and is thought to be the only common subunit required by all three RNA polymerases.
Concentration: 100 µg/12,5 µl
Description: Monoclonal antibody raised in mouse against the amino-terminal domain of human TBP (TATA box binding protein).
Format: Aqueous buffer solution
Formulation: PBS containing 0.05% azide
Immunogen: recombinant protein
Purification: Purified by ammonium sulphate precipitation followed by dialysis
Storage Stability: Store at -80°C for up to 2 years. Centrifuge after first thaw to maximize product recovery. Aliquot to avoid repeated freeze/thaw cycles. Aliquots may be stored at -20°C for at least one month.
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
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