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Epigenetics and Nuclear Signaling Histones Variants

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

  • Datasheet
  • SDS
Reviews (21)Q&A (20)References (120)

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

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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 corresponding to Human Histone H2A.Z aa 100 to the C-terminus conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab11681)

  • 常规说明

    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.

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)
  • Immunizing Peptide (Blocking)

    • Human Histone H2A.Z peptide (ab11681)
  • 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

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.

应用 Ab评论 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.

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

    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.

  • 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

    • Datasheet
    • SDS
  • 文献 (120)

    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 120 文献中.

    • 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
    View all Publications for this product

    客户评价及客户问答

    Show All 评价 Q&A
    提交评价 提交问题

    1-10 of 41 Abreviews or Q&A

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

    Excellent
    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

    Abcam user community

    Verified customer

    提交于 Dec 12 2011

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

    Inconclusive
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Arabidopsis thaliana Tissue lysate - nuclear (50 6-day old Arabidopsis seedlings)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    0.4 µg
    Specification
    50 6-day old Arabidopsis seedlings
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 28°C
    Read More

    Abcam user community

    Verified customer

    提交于 Nov 20 2020

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

    Average
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (mouse Oli-neu cells)
    Permeabilization
    Yes - 0.05% triton-x100
    Specification
    mouse Oli-neu cells
    Blocking step
    Vetor Blocker as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 23°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    提交于 Dec 07 2019

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

    Inconclusive
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Saccharomyces cerevisiae Cell lysate - whole cell (WT and htz1 delta (H2A.Z deletion ) cells)
    Gel Running Conditions
    Non-reduced Denaturing (15% gel)
    Loading amount
    1e+007 cells
    Specification
    WT and htz1 delta (H2A.Z deletion ) cells
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 30°C
    Read More

    Daisuke Takahashi

    Verified customer

    提交于 Aug 13 2018

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

    Below Average
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HeLa)
    Permeabilization
    Yes - 0.5% Triton 5 minutes
    Specification
    HeLa
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    提交于 Jan 25 2018

    Immunohistochemistry (Resin sections) abreview for Anti-Histone H2A.Z antibody - ChIP Grade

    Average
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Resin sections)
    Sample
    Astatotilapia burtoni Tissue sections (Brain preopticarea embedded in LRWhite)
    Specification
    Brain preopticarea embedded in LRWhite
    Read More

    Sebastian Alvarado

    Verified customer

    提交于 May 03 2016

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

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    0.5 µg
    Gel Running Conditions
    Reduced Denaturing (4-20% TGX (BioRad))
    Sample
    Human Purified protein (Recombinant octamers & purified K562 cell histones)
    Specification
    Recombinant octamers & purified K562 cell histones
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 20°C
    Read More

    Dr. Ragnhild Eskeland

    Verified customer

    提交于 Jan 27 2015

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

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    ChIP
    Detection step
    Other
    Sample
    Mouse Cell lysate - whole cell (bone marrow derived mouse macrophages)
    Specification
    bone marrow derived mouse macrophages
    Negative control
    IgG
    Type
    Cross-linking (X-ChIP)
    Duration of cross-linking step: 10 minute(s) and 0 second(s)
    Specification of the cross-linking agent: formaldehyde 1% final
    Positive control
    Pu.1 antibody
    Read More

    Dr. Silvia Bonifacio

    Verified customer

    提交于 Oct 24 2014

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

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    20 µg
    Gel Running Conditions
    Reduced Denaturing (15%)
    Sample
    Arabidopsis thaliana Tissue lysate - nuclear (7dps seedlings)
    Specification
    7dps seedlings
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Read More

    Abcam user community

    Verified customer

    提交于 Sep 25 2014

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

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    ChIP
    Detection step
    Real-time PCR
    Sample
    Mouse Cell lysate - whole cell (Embryonic Stem Cells)
    Specification
    Embryonic Stem Cells
    Type
    Cross-linking (X-ChIP)
    Duration of cross-linking step: 10 minute(s) and 0 second(s)
    Specification of the cross-linking agent: formaldehyde
    Read More

    Mr. Dan Stummer

    Verified customer

    提交于 Mar 29 2014

    1-10 of 41 Abreviews or Q&A

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