Anti-Histone H2A.Z 抗体 - ChIP Grade (ab4174)
Key features and details
- Rabbit polyclonal to Histone H2A.Z - ChIP Grade
- Suitable for: ICC/IF, ChIP, WB
- Reacts with: Mouse, Rat, Cow, Human
- Isotype: IgG
リコンビナント抗体で、ロット間での高い再現性を実現
- 異なるロット間での安定した再現性
- 容易なスケールアップ
- 評価試験による特異性の確認済み
- 倫理基準に準拠 - アニマル・フリーの生産
製品の概要
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製品名
Anti-Histone H2A.Z antibody - ChIP Grade
Histone H2A.Z 一次抗体 製品一覧 -
製品の詳細
Rabbit polyclonal to Histone H2A.Z - ChIP Grade -
由来種
Rabbit -
特異性
From Jan 2024, QC testing of replenishment batches of this polyclonal changed. All tested and expected application and reactive species combinations are still covered by our Abcam product promise. However, we no longer test all applications. For more information on a specific batch, please contact our Scientific Support who will be happy to help.
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アプリケーション
適用あり: ICC/IF, ChIP, WBmore details -
種交差性
交差種: Mouse, Rat, Cow, Human
交差が予測される動物種: Sheep, Xenopus laevis, Arabidopsis thaliana, Zebrafish -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
(Peptide available asab11681) -
ポジティブ・コントロール
- ICC/IF: HepG2
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特記事項
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
製品の特性
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製品の状態
Liquid -
保存方法
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
バッファー
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help. -
Concentration information loading...
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精製度
Immunogen affinity purified -
ポリ/モノ
ポリクローナル -
アイソタイプ
IgG -
研究分野
関連製品
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ChIP Related Products
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
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Related Products
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab4174の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
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ICC/IF | (3) |
1/1000.
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ChIP | (9) |
Use at an assay dependent concentration.
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WB | (14) |
1/1000. Detects a band of approximately 14 kDa (predicted molecular weight: 13.4 kDa).
Abcam recommends using BSA as the blocking agent. |
特記事項 |
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ICC/IF
1/1000. |
ChIP
Use at an assay dependent concentration. |
WB
1/1000. Detects a band of approximately 14 kDa (predicted molecular weight: 13.4 kDa). Abcam recommends using BSA as the blocking agent. |
ターゲット情報
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機能
Variant histone H2A which replaces conventional H2A in a subset of nucleosomes. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling. May be involved in the formation of constitutive heterochromatin. May be required for chromosome segregation during cell division. -
配列類似性
Belongs to the histone H2A family. -
翻訳後修飾
Monoubiquitination of Lys-122 gives a specific tag for epigenetic transcriptional repression.
Acetylated on Lys-5, Lys-8 and Lys-12 during interphase. Acetylation disappears at mitosis.
Monomethylated on Lys-5 and Lys-8 by SETD6. SETD6 predominantly methylates Lys-8, lys-5 being a possible secondary site.
Not phosphorylated. -
細胞内局在
Nucleus. Chromosome. - Information by UniProt
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参照データベース
- Entrez Gene: 287016 Cow
- Entrez Gene: 3015 Human
- Entrez Gene: 51788 Mouse
- Entrez Gene: 58940 Rat
- Omim: 142763 Human
- SwissProt: P0C0S4 Cow
- SwissProt: P0C0S5 Human
- SwissProt: P0C0S6 Mouse
see all -
別名
- H2A histone family member Z antibody
- H2A.z antibody
- H2A/z antibody
see all
画像
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Changes in nucleosome occupancy upon LPS induction at a putative distal enhancer and promoter of IL1A, kinetics of nucleosome removal, and changes in histone modifications.
Chromation from mouse bone marrow derived macrophages. ChIP experiments with antibodies against H3 (dark blue bars), H2A.Z (light blue, (ab4174 at 4µg), H3K4me1 (green, ab8895 at 1 µg), H3K4me3 (yellow, ab8580 at 1 µg) and H3K27ac (red, ab4729 at 1 µg). For these experiments cross-linked chromatin was lightly digested with MNase before incubation with the respective antibodies to increase resolution of the ChIP signal and the data was normalized to a region in the ORF of RPL4. Changes upon LPS induction in histone binding and histone modifications at the enhancers and promoters of IL12B and IL1A as well as at a control region in the GAPDH pseudo gene are shown as fold over levels found before induction. For H3K27ac the changes 1.5 h after LPS induction, and for all other histone variants and modifications the changes after 3 h of induction are shown. The error bars show the SEM of at least 3 independent experiments. Statistical significance of the changes in H3K4me3 and H3K27ac upon LPS induction compared to levels found prior to induction determined by Student's T-tests is indicated (*P<0.05; **P<0.01).
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Chromatin was prepared from HeLa (Human epithelial cell line from cervix adenocarcinoma) cells according to the Abcam X-ChIP protocol.
Cells were fixed with formaldehyde for 10 min. The ChIP was performed with 25 µg of chromatin, 2 µg of ab4174 (blue), and 20 µl of Protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci, Sybr green approach for heterochromatic loci). Primers and probes are located in the first kb of the transcribed region.
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ab4174 staining Histone H2A.Z in HepG2 cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab4174 at 1µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
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Lanes 1-2 : Anti-Histone H2A.Z antibody - ChIP Grade (ab4174) at 1/500 dilution
Lanes 3-4 : Anti-Histone H2A.Z antibody - ChIP Grade (ab4174) at 1/1000 dilution
Lanes 1 & 3 : Calf thymus histone lysate
Lanes 2 & 4 : Calf thymus histone lysate with Human Histone H2A.Z peptide (ab11681) at 1 µg/ml
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/5000 dilution
Performed under reducing conditions.
Predicted band size: 13.4 kDa -
Staining of Histone H2A.Z in mouse embryonic cells. The fixation is 2% PFA, and permeabilization is PBS 0.5% triton BSA. The dilution used was 1 in 100 (but it could be used at 1/200 to 1/300).
Red = H2A.Z
Blue = toto3 for the DNA -
All lanes : Anti-Histone H2A.Z antibody - ChIP Grade (ab4174) at 1 µg/ml
Lane 1 : Calf Thymus Histone Preparation Nuclear Lysate at 0.5 µg
Lane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg
Lane 3 : NIH/3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 10 µg
Lane 4 : PC-12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate at 10 µg
Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) (ab65484) at 1/3000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 13.4 kDa
Exposure time: 3 minutes -
All lanes : Anti-Histone H2A.Z antibody - ChIP Grade (ab4174) at 1/1000 dilution
Lane 1 : Native recombinant octamers K562 (Human chronic myelogenous leukemia cell line from bone marrow) cells at 3 µg
Lane 2 : Native recombinant octamers K562 cells at 1.5 µg
Lane 3 : Recombinant Human octamers containing H2A at 1 µg
Lane 4 : Recombinant Human octamers containing H2A at 0.5 µg
Lane 5 : Recombinant Human octamers containing H2A.Z.2.1 at 0.5 µg
Lane 6 : Recombinant Human octamers containing H2A.Z.1 at 0.5 µg
Secondary
All lanes : HRP-conjugated donkey anti-rabbit IgG polyclonal at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 13.4 kDa
Exposure time: 5 minutesBlocked with 3% BSA for 1 hour at 20°C.
Primary incubation in TBS tween + 3% BSA at 20°C for 1 hour.
プロトコール
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (142)
ab4174 は 142 報の論文で使用されています。
- Yang Y et al. HIRA complex presets transcriptional potential through coordinating depositions of the histone variants H3.3 and H2A.Z on the poised genes in mESCs. Nucleic Acids Res 50:191-206 (2022). PubMed: 34893908
- Shi Y et al. Critical role of Znhit1 for postnatal heart function and vacuolar cardiomyopathy. JCI Insight 7:N/A (2022). PubMed: 35167494
- Sump B et al. Mitotically heritable, RNA polymerase II-independent H3K4 dimethylation stimulates INO1 transcriptional memory. Elife 11:N/A (2022). PubMed: 35579426
- Wei W et al. Chromatin remodeler Znhit1 controls bone morphogenetic protein signaling in embryonic lung tissue branching. J Biol Chem 298:102490 (2022). PubMed: 36115458
- Reske JJ et al. ARID1A-dependent maintenance of H3.3 is required for repressive CHD4-ZMYND8 chromatin interactions at super-enhancers. BMC Biol 20:209 (2022). PubMed: 36153585