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Epigenetics and Nuclear Signaling Histones Variants

Anti-Histone H2A.Z抗体- ChIP Grade (ab4174)

  • Datasheet
  • SDS
Reviews (30)Q&A (20)References (129)

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ChIP - Anti-Histone H2A.Z antibody - ChIP Grade (ab4174)
  • ChIP - Anti-Histone H2A.Z antibody - ChIP Grade (ab4174)
  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H2A.Z antibody - ChIP Grade (ab4174)
  • Western blot - Anti-Histone H2A.Z antibody - ChIP Grade (ab4174)
  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H2A.Z antibody - ChIP Grade (ab4174)
  • Western blot - Anti-Histone H2A.Z antibody - ChIP Grade (ab4174)
  • Western blot - Anti-Histone H2A.Z antibody - 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

选择批间可重复性更高的重组抗体

Product image
Anti-Histone H2A.Z antibody [EPR6171(2)(B)] - ChIP Grade (ab150402)
  • 研究可靠 —— 各批次间结果一致且可重复
  • 长期批量供应 —— 采用重组技术,可实现快速生产
  • 首次实验即可成功 —— 经过大量验证确认了特异性
  • 符合伦理标准 —— 产品不含动物成分

概述

  • 产品名称

    Anti-Histone H2A.Z抗体- ChIP Grade
    参阅全部 Histone H2A.Z 一抗
  • 描述

    兔多克隆抗体to Histone H2A.Z - ChIP Grade
  • 宿主

    Rabbit
  • 经测试应用

    适用于: ICC/IF, ChIP, WBmore details
  • 种属反应性

    与反应: Mouse, Rat, Cow, Human
    预测可用于: Sheep, Xenopus laevis, Arabidopsis thaliana, Zebrafish
  • 免疫原

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
    (Peptide available as ab11681)

  • 阳性对照

    • ICC/IF: HepG2
  • 常规说明

    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

性能

  • 形式

    Liquid
  • 存放说明

    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.
  • 存储溶液

    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...
  • 纯度

    Immunogen affinity purified
  • 克隆

    多克隆
  • 同种型

    IgG
  • 研究领域

    • Epigenetics and Nuclear Signaling
    • Histones
    • Variants
    • Epigenetics and Nuclear Signaling
    • ChIP assays
    • ChIP antibodies

相关产品

  • ChIP Related Products

    • Rabbit Anti-Mouse IgG H&L (ab46540)
    • ChIP Kit (ab500)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Positive Controls

    • Recombinant Human Histone H2A.Z protein (ab114184)
  • Recombinant Protein

    • Recombinant Human Histone H2A.Z protein (denatured) (ab177637)
  • Related Products

    • Prestained Protein Ladder – Broad molecular weight (10-245 kDa) (ab116028)

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab4174于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
ICC/IF (3)
1/1000.
ChIP (9)
Use at an assay dependent concentration.
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.

说明
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.

靶标

  • 功能

    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.
  • 序列相似性

    Belongs to the histone H2A family.
  • 翻译后修饰

    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.
  • 细胞定位

    Nucleus. Chromosome.
  • Target information above from: UniProt accession P0C0S5 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 数据库链接

    • 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
    • SwissProt: P0C0S7 Rat
    • SwissProt: Q6YNC8 Sheep
    • Unigene: 119192 Human
    • Unigene: 117541 Mouse
    • Unigene: 372513 Mouse
    • Unigene: 465508 Mouse
    • Unigene: 112573 Rat
    see all
  • 别名

    • H2A histone family member Z antibody
    • H2A.z antibody
    • H2A/z antibody
    • H2afz antibody
    • H2AZ antibody
    • H2AZ_HUMAN antibody
    • Histone H2A.Z antibody
    • MGC117173 antibody
    see all

图片

  • ChIP - Anti-Histone H2A.Z antibody - ChIP Grade (ab4174)
    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).

  • ChIP - Anti-Histone H2A.Z antibody - ChIP Grade (ab4174)
    ChIP - Anti-Histone H2A.Z antibody - ChIP Grade (ab4174)

    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.

  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H2A.Z antibody - ChIP Grade (ab4174)
    Immunocytochemistry/ Immunofluorescence - Anti-Histone H2A.Z antibody - ChIP Grade (ab4174)

    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.

  • Western blot - Anti-Histone H2A.Z antibody - ChIP Grade (ab4174)
    Western blot - Anti-Histone H2A.Z antibody - ChIP Grade (ab4174)
    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

  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H2A.Z antibody - ChIP Grade (ab4174)
    Immunocytochemistry/ Immunofluorescence - Anti-Histone H2A.Z antibody - ChIP Grade (ab4174)

    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

  • Western blot - Anti-Histone H2A.Z antibody - ChIP Grade (ab4174)
    Western blot - Anti-Histone H2A.Z antibody - ChIP Grade (ab4174)
    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
  • Western blot - Anti-Histone H2A.Z antibody - ChIP Grade (ab4174)
    Western blot - Anti-Histone H2A.Z antibody - ChIP Grade (ab4174)This image is courtesy of an Abreview submitted by Ragnhild Eskeland
    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 minutes


    Blocked with 3% BSA for 1 hour at 20°C.

    Primary incubation in TBS tween + 3% BSA at 20°C for 1 hour.

    See Abreview

实验方案

  • ChIP protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

数据表及文件

  • SDS download

  • Datasheet download

    Download

文献 (129)

发表研究结果有使用 ab4174?请让我们知道,以便我们可以引用本数据表中的参考文章。

ab4174 被引用在 129 文献中.

  • Perampalam P  et al. Disrupting the DREAM transcriptional repressor complex induces apolipoprotein overexpression and systemic amyloidosis in mice. J Clin Invest 131:N/A (2021). PubMed: 33444292
  • Schachner LF  et al. Decoding the protein composition of whole nucleosomes with Nuc-MS. Nat Methods 18:303-308 (2021). PubMed: 33589837
  • Raja DA  et al. Histone variant dictates fate biasing of neural crest cells to melanocyte lineage. Development 147:N/A (2020). PubMed: 32098766
  • Tong M  et al. The Evening Complex Establishes Repressive Chromatin Domains Via H2A.Z Deposition. Plant Physiol 182:612-625 (2020). PubMed: 31712305
  • 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
View all Publications for this product

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Western blot abreview for Anti-Histone H2A.Z antibody - ChIP Grade

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Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Mouse Cell lysate - nuclear (Mouse embryonic fibroblasts)
Loading amount
25 µg
Specification
Mouse embryonic fibroblasts
Gel Running Conditions
Reduced Denaturing (4-20% Tris-Gly gel)
Blocking step
LI-COR® Odyssey® Blocking Buffer as blocking agent for 45 minute(s) · Concentration: 50% · Temperature: RT°C
Read More
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Western blot abreview for Anti-Histone H2A.Z antibody - ChIP Grade

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Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (Lung cell line)
Gel Running Conditions
Reduced Denaturing (12)
Loading amount
60 µg
Specification
Lung cell line
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
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提交于 Dec 01 2021

ChIP abreview for Anti-Histone H2A.Z antibody - ChIP Grade

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abreview image
Application
ChIP
Sample
Mouse Cell lysate - whole cell (Cell line)
Specification
Cell line
Detection step
Real-time PCR
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: Formaldehyde
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提交于 Nov 16 2021

Immunocytochemistry/ Immunofluorescence abreview for Anti-Histone H2A.Z antibody - ChIP Grade

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Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (CRC cell line)
Permeabilization
Yes - Tween
Specification
CRC cell line
Blocking step
Serum as blocking agent for 15 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
Acetone
Read More
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提交于 Nov 13 2021

Western blot abreview for Anti-Histone H2A.Z antibody - ChIP Grade

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Application
Western blot
Sample
Human Cell lysate - nuclear (T-ALL cell line)
Gel Running Conditions
Reduced Denaturing (12%)
Loading amount
30 µg
Specification
T-ALL cell line
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
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提交于 Nov 10 2021

ChIP on chip abreview for Anti-Histone H2A.Z antibody - ChIP Grade

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ChIP on chip
Sample
Mouse Cell lysate - whole cell (cell line)
Specification
cell line
Detection step
Microarray
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Read More
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提交于 Oct 28 2021

ChIP abreview for Anti-Histone H2A.Z antibody - ChIP Grade

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ChIP
Sample
Mouse Cell lysate - whole cell (Cell line)
Specification
Cell line
Detection step
Real-time PCR
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: 1% Formaldehyde
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提交于 Oct 12 2021

Western blot abreview for Anti-Histone H2A.Z antibody - ChIP Grade

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Application
Western blot
Sample
Mouse Cell lysate - nuclear (Cell line)
Gel Running Conditions
Reduced Denaturing (12%)
Loading amount
97 µg
Specification
Cell line
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
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ChIP on chip abreview for Anti-Histone H2A.Z antibody - ChIP Grade

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ChIP on chip
Sample
Mouse Cell lysate - whole cell (cell line)
Specification
cell line
Detection step
Microarray
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: Formaldehyde
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Western blot abreview for Anti-Histone H2A.Z antibody - ChIP Grade

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Western blot
Sample
Mouse Cell lysate - whole cell (Cell line)
Gel Running Conditions
Reduced Denaturing (12%)
Loading amount
34 µg
Specification
Cell line
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Read More
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