AF9 (UniProtKB/Swiss-Prot entry P42568) is a component of the super elongation complex which is required to increase the catalytic rate of RNA polymerase II transcription by suppressing transient pausing by the polymerase at multiple sites along the DNA.
AF9 Antibody - ChIP-seq Grade (sample size)
Other names: MLLT3, YEATS3
Polyclonal antibody raised in rabbit against human AF9 (Super Elongation Complex Subunit) using a KLH conjugated synthetic peptide containing a sequence from the central region of the protein.
| Lot | A1474-001 |
|---|---|
| Concentration | Not determined |
| Species reactivity | Human: positive. Other species: not tested. |
| Type | Polyclonal ChIP grade, ChIP-seq grade |
| Purity | Whole antiserum from rabbit 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/amount | References |
|---|---|---|
| ChIP/ChIP-seq * | 2 µl/ChIP | Fig 1, 2 |
| ELISA | 1:100 - 1:500 | Fig 3 |
*Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-10 µl per IP.
- Validation Data
Figure 1. ChIP results obtained with the Diagenode antibody directed against AF9
ChIP was performed on THP-1 cells using the Diagenode antibody against AF9 (Cat. No. C15310266). Sheared chromatin from 1 million cells and 2 μl of antibody were used per ChIP experiment. QPCR was performed using primers specific for the indicated genes. Figure 1 shows the recovery (the relative amount of immunoprecipitated DNA compared to input DNA).
Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against AF9
ChIP was performed as described above. The IP’d DNA of 5 ChIP’s was pooled and 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 400 kb region of chromosome 6 containing the PHIP positive control gene (fig 2A and B), and in 3 genomic regions surrounding the CDKN2C gene, the JMJD1C gene and HOX cluster on chromosome 1, 10 and 7, respectively (fig 2C, D and E).
Figure 3. Determination of the antibody titer
To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against AF9 (Cat. No. C15310266). The plates were coated with the peptide 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 1:8,000. - 出版物
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