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
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 -
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 corresponding to Human Histone H2A.Z aa 100 to the C-terminus conjugated to keyhole limpet haemocyanin.
(Peptide available asab11681) -
特記事項
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
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, as well as 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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Immunizing Peptide (Blocking)
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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
Our Abpromise guarantee covers the use of ab4174 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
アプリケーション | Abreviews | 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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ChIP - Anti-Histone H2A.Z antibody - ChIP Grade (ab4174)Gjidoda et al PLoS One. 2014 Apr 4;9(4):e93971. doi: 10.1371/journal.pone.0093971. eCollection 2014. Fig 2. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/
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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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 withHuman 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 -
ICC/IF image of ab4174 stained HepG2 (Human liver hepatocellular carcinoma cell line) cells.
The cells were fixed with 100% methanol (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilize the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab4174, 5 µg/ml) overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti-rabbit IgG - H&L, pre-adsorbed (ab96899) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
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Western blot - Anti-Histone H2A.Z antibody - ChIP Grade (ab4174)This image is courtesy of an Abreview submitted by Ragnhild EskelandAll 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
参考文献 (120)
ab4174 は 120 報の論文で使用されています。
- Wen Z et al. Histone variant H2A.Z regulates nucleosome unwrapping and CTCF binding in mouse ES cells. Nucleic Acids Res 48:5939-5952 (2020). PubMed: 32392318
- Nakabayashi Y et al. An improved functional analysis of linker-mediated complex (iFALC) strategy. Biochem Biophys Res Commun 526:1164-1169 (2020). PubMed: 32327258
- Lamaa A et al. Integrated analysis of H2A.Z isoforms function reveals a complex interplay in gene regulation. Elife 9:N/A (2020). PubMed: 32109204
- Greenberg RS et al. Single Amino Acid Change Underlies Distinct Roles of H2A.Z Subtypes in Human Syndrome. Cell 178:1421-1436.e24 (2019). PubMed: 31491386
- Yamada T et al. Sensory experience remodels genome architecture in neural circuit to drive motor learning. Nature 569:708-713 (2019). PubMed: 31068695