HDAC6 (UniProt/Swiss-Prot entry Q9UBN7) belongs to class II of the histone deacetylase family. These enzymes catalyse the deacetylation of lysine residues on the N-terminal part of the core histones (H2A, H2B, H3 and H4). Histone deacetylation plays an important role in transcriptional regulation, cell cycle progression and developmental events. HDAC6 also plays a key role in the degradation of misfolded proteins and in microtubuledependent cell motility via deacetylation of tubulin and is involved in the MTA1-mediated epigenetic regulation of ESR1 expression in breast cancer.
HDAC6 Antibody - ChIP-seq Grade (sample size)
Other names: HD6, PPP1R90, CPBHM, JM21
Polyclonal antibody raised in rabbit against human HDAC6 (Histone deacetylase 6), using two KLH-conjugated synthetic peptides from the N-terminal and the central part of the protein, respectively.
| Lot | A2850P |
|---|---|
| Concentration | 0.5 µg/µl |
| Species reactivity | Human: positive. Other species: not tested. |
| Type | Polyclonal, ChIP grade, ChIP-seq grade |
| Purity | Affinity purified polyclonal antibody. |
| Host | Rabbit |
| Storage Conditions | Store at -20°C; for long storage, store at -80°C. Avoid multiple freeze-thaw cycles. |
| Storage Buffer | PBS containing 0.05% azide. |
| Precautions | This product is for research use only. Not for use in diagnostic or therapeutic procedures. |
| Applications | Suggested dilution | References |
|---|---|---|
| ChIP/ChIP-seq* | 3 µg/ChIP | Fig 1, 2 |
| ELISA | 1:1,000 | Fig 3 |
| Western Blotting | Not recommended |
*Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1 - 10 μg per IP.
- Validation data
Figure 1. ChIP results obtained with the Diagenode antibody directed against HDAC6
ChIP was performed with the Diagenode antibody against HDAC6 (cat. No. C15410362) on sheared chromatin from 4,000,000 HeLa cells with the iDeal ChIP-seq kit for TF’s. An antibody titration consisting of 0.5, 1, 3 and 6 µg per ChIP experiment was analysed. IgG (1 µg/IP) was used as negative IP control. QPCR was performed with primers specific for the GAPDH and EIF4A2 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 HDAC6
ChIP was performed on sheared chromatin from 4,000,000 HeLa cells using 3 µg of the Diagenode antibody against HDAC6 (cat. No. C15410362) 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 peak distribution along the complete sequence and a 300 kb region of chromosome 1 (figure 2A and B) and in two genomic regions surrounding the EIF2S3 and GAPDH positive control genes, respectively (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 HDAC6 (cat. No. C15410362). 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 1:11,400. - 出版物
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