| ChIP-seq 検出方法（ChIP-seq）としてハイスループットシーケンシングと結合したクロマチン免疫沈降（ChIP）は、今やエピゲノム研究、すなわちゲノム規模でのタンパク質-DNA相互作用の研究において主要な方法の1つされています。また、この技術は現在、細胞分化、腫瘍抑制遺伝子のサイレンシング、および遺伝子発現に対するヒストン修飾の効果を含む様々なライフサイエンス分野で使用されています。 ChIP-seqワークフロー クロマチン調製 : DNAへのヒストンまたは転... Read more |
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H2A.Z monoclonal antibody
Catalog Number
Format
Price
Polyclonal antibody raised in rabbit against histone variant H2A.Z, using a KLH-conjugated synthetic peptide containing a sequence from the C-terminal part of the protein.
| Lot | 001D |
|---|---|
| Concentration | 1 µg/µl |
| Species reactivity | Human: positive. Other species: not tested |
| Type | Monoclonal 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 50% glycerol, 1% BSA and 0.09% azide |
| Precautions | This product is for research use only. Not for use in diagnostic or therapeutic procedures. |
| Applications | Suggested dilution | References |
|---|---|---|
| ChIP/ChIP-seq * | 0.5 µg per IP | Fig 1, 2 |
| Western Blotting | 1:200 | Fig 3 |
| Immunofluorescence | 1:100 | Fig 4 |
*Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 0.5 - 5 µg per IP.
- Validation data
Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H2A.Z
ChIP was performed with the Diagenode antibody against H2A.Z (cat. No. C15210021) on sheared chromatin from 500,000 HeLaS3 cells using the “iDeal ChIP-seq” kit (cat. No. C01010051). A titration of the antibody consisting of 0.5, 1, and 2.5 µg per ChIP experiment was analysed. IgG (1 µg/IP) was used as negative IP control. Quantitative PCR was performed with primers for the EIF2S3 and EIF4A2 promoters, used as positive controls, and for the MYOD1 and MYT1 genes, used as negative controls. The graph shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).A.
B.
C.
D.
Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H2A.Z
ChIP was performed on sheared chromatin from 500,000 HeLaS3 cells using 0.5 µg of the Diagenode antibody against H2A.Z (cat. No. C15210021) as described above. The IP'd DNA was subsequently analysed on an Illumina NovaSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the H2A.Z signal along the complete sequence and 1.5 Mb region of the human X-chromosome (figure 2A and B), and in two genomic regions surrounding the EIF2S3 and EIF4A2 positive controls (figure 2C and D).
Figure 3. Western blot analysis using the Diagenode monoclonal antibody directed against H2A.Z
Western blot was performed on whole cell extracts (40 µg) from HeLa cells using the Diagenode antibody against H2A.Z (cat. No. C15210021). The antibody was diluted 1:200 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is shown on the right, the marker (in kDa) is shown on the left.
Figure 4. Immunofluorescence using the Diagenode monoclonal antibody directed against H2A.Z
HeLa cells were stained with the Diagenode antibody against H2A.Z (cat. No. C15210021, red) diluted 1:100. Actin was stained with fluorescein phalladoin (green). - 出版物
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