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

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
  • 经测试应用适用于: ICC/IF, ChIP, IP, CHIPseq, WBmore details
  • 种属反应性
    与反应: Mouse, Rat, Rabbit, Chicken, Cow, Human, Arabidopsis thaliana, Zebrafish
    预测可用于: Sheep, Xenopus laevis
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 100 to the C-terminus of Human Histone H2A.Z.

    (Peptide available as ab11681.)

  • 阳性对照
    • This antibody gave a positive control in the following lysate: Calf thymus histone lysate; HeLa whole cell. This antibody gave a positive control in the following mouse lysates: NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell MEF1 (Mouse embryonic fibroblast cell line) Whole Cell This antibody gave a positive control in the following rat lysate: PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell

性能

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.
IP Use at an assay dependent concentration.
CHIPseq Use at an assay dependent concentration. PubMed: 22196736
WB 1/1000. Detects a band of approximately 14 kDa (predicted molecular weight: 13.4 kDa).

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.
  • Information by UniProt
  • 数据库链接
  • 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

Anti-Histone H2A.Z antibody - ChIP Grade 图像

  • Chromatin was prepared from Hela 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.



  • Performed under reducing conditions.

    Predicted band size : 13.4 kDa
  • The staining has been done on 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 to1/300)

    Red  = H2A.Z
    Blue  = toto3 for the DNA


  • Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 13.4 kDa


    Exposure time : 3 minutes

    Lane 1 Primary: ab4174, 1 ug/ml
    Sample: Calf Thymus Histone Preparation Nuclear Lysate, 0.5 ug
    Secondary: ab65484, 1/3000 dilution

    Lane 2 Primary: ab4174, 1 ug/ml
    Sample: HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate, 10 ug
    Secondary: ab65484, 1/3000 dilution

    Lane 3 Primary: ab4174, 1 ug/ml
    Sample: NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate, 10 ug
    Secondary: ab65484, 1/3000 dilution

    Lane 4 Primary: ab4174, 1 ug/ml
    Sample: PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate , 10 ug
    Secondary: ab65484, 1/3000 dilution

  • This image was kindly supplied as part of the review submitted by Dr Geza Fejes-Toth.
  • ICC/IF image of ab4174 stained HepG2 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab4174, 5µg/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.


  • Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 13.4 kDa


    Exposure time : 5 minutes

    This image is courtesy of an Abreview submitted by Ragnhild Eskeland

    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

Anti-Histone H2A.Z antibody - ChIP Grade (ab4174)参考文献

This product has been referenced in:
  • Saito T  et al. Histone modification and signalling cascade of the dormancy-associated MADS-box gene, PpMADS13-1, in Japanese pear (Pyrus pyrifolia) during endodormancy. Plant Cell Environ 38:1157-66 (2015). Read more (PubMed: 25311427) »
  • Yang X  et al. Gene body methylation can alter gene expression and is a therapeutic target in cancer. Cancer Cell 26:577-90 (2014). ChIP ; Human . Read more (PubMed: 25263941) »

See all 46 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Abcam user community

Verified customer

提交于 Dec 12 2011

Application Immunohistochemistry (Resin sections)
Sample Astatotilapia burtoni Tissue sections (Brain preopticarea embedded in LRWhite)
Specification Brain preopticarea embedded in LRWhite
Username

Dr. Sebastian Alvarado

Verified customer

提交于 May 03 2016

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
Username

Dr. Ragnhild Eskeland

Verified customer

提交于 Jan 27 2015

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
Username

Dr. Silvia Bonifacio

Verified customer

提交于 Oct 24 2014

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
Username

Abcam user community

Verified customer

提交于 Sep 25 2014

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
Username

Mr. Dan Stummer

Verified customer

提交于 Mar 29 2014

Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing
Sample Mouse Cell lysate - whole cell (Murine embryonic stem cells)
Specification Murine embryonic stem cells
Treatment vehicle control (EtOH) or all-trans retinoic acid (RA)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Mr. Dan Stummer

Verified customer

提交于 Mar 28 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application ChIP
Sample Human Cell lysate - nuclear (haematopoietic)
Specification haematopoietic
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: formaldehyde
Detection step Real-time PCR
Positive control PU.1
Negative control IVL
Username

Abcam user community

Verified customer

提交于 Dec 26 2012

Thank you for your reply. I am very pleased to hear you would like to accept our offer and test ab4174 in Arabidopsis.

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I am sorry to confirm that regrettably we have not yet received feedback from these customers on the use of ab4174 on Arabidopsis in ChIP.

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