YY1 (UniProtKB/Swiss-Prot entry P25490) is a ubiquitously distributed transcription factor which can both activate or repress a large number of cellular and viral genes by binding to sites overlapping the transcription start site. Whether it activates or represses transcription depends upon the context in which it binds. YY1 is thought to direct histone deacetylases and histone acetyltransferases to the promoters of it’s target genes in order to activate or repress transcription, thus implicating histone modification in the function of YY1.
YY1 Antibody - ChIP-seq Grade (sample size)
Other names: INO80S, NF-E1, YY-1, Yin-Yang-1, UCRBP
Polyclonal antibody raised in rabbit against human Transcription factor YY1, using two synthetic peptides containing a sequence from the central part and the C-terminus of the protein, respectively.
| Lot | A2649-0040 |
|---|---|
| Concentration | 2.3 μg/μl |
| Species reactivity | Human: positive. Other species: not tested. |
| Type | Polyclonal |
| Purity | Affinity purified polyclonal antibody in PBS containing 0.05% azide. |
| Host | Rabbit |
| Storage Conditions | Store at -20°C; for long storage, store at -80°C. Avoid multiple freeze-thaw cycles. |
| Precautions | This product is for research use only. Not for use in diagnostic or therapeutic procedures. |
| Applications | Suggested dilution | References |
|---|---|---|
| ChIP/ChIP-seq * | 1 μg/ChIP | Fig 1, 2 |
| ELISA | 1:10,000 - 1:100,000 | Fig 3 |
| WB | 1:1,000 | Fig 4 |
* Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-5 μg per IP.
- Validation data
Figure 1. ChIP results obtained with the Diagenode antibody directed against YY1
ChIP assays were performed using K562 cells, the Diagenode antibody against YY1 (Cat. No. C15410345) and optimized PCR 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 consisting of 1, 2, 5 and 10 μg of antibody per ChIP experiment was analyzed. IgG (2 μg/IP) was used as a negative IP control. Quantitative PCR was performed with primers for the promoters of the FBXL18 and EIF2S3 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 YY1
ChIP was performed with 1 μg of the Diagenode antibody against YY1 (Cat. No. C15410345) on sheared chromatin from 4,000,000 K562 cells using the “iDeal ChIP-seq” kit as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq 4000. 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 signal distribution along the complete sequence and a 1 Mb region of the human X-chromosome (figures 2A and B), and in two genomic regions surrounding the FBXL18 and EIF2S3 positive control genes (figure 2C and D).
Figure 3. Determination of the antibody titer
To determine the titer of the antibody, an ELISA was performed using a serial dilution of Diagenode antibody directed against YY1 (Cat. No. C15410345). The plates were coated with the peptides used for immunization of the rabbit. By plotting the absorbance against the antibody dilution (Figure 3), the titer of the antibody was estimated to be >1:1,000,000.
Figure 4. Western blot analysis using the Diagenode antibody directed against YY1
Whole cell extracts from K562 cells were analysed by Western blot using the Diagenode antibody against YY1 (Cat. No. C15410345) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left. - 出版物
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