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

Overview

  • Product name
    Anti-Histone H2A.Z antibody - ChIP Grade
    See all Histone H2A.Z primary antibodies
  • Description
    Rabbit polyclonal to Histone H2A.Z - ChIP Grade
  • Tested applications
    Suitable for: ICC/IF, ChIP, IP, CHIPseq, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Rabbit, Chicken, Cow, Human, Arabidopsis thaliana, Zebrafish
    Predicted to work with: Sheep, Xenopus laevis
  • Immunogen

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

    (Peptide available as ab11681.)

  • Positive control
    • 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

Properties

Applications

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.

Application Abreviews Notes
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).

Target

  • Function
    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.
  • Sequence similarities
    Belongs to the histone H2A family.
  • Post-translational
    modifications
    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.
  • Cellular localization
    Nucleus. Chromosome.
  • Information by UniProt
  • Database links
  • Alternative names
    • 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 images

  • 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

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

This product has been referenced in:
  • Barski A  et al. Rapid Recall Ability of Memory T cells is Encoded in their Epigenome. Sci Rep 7:39785 (2017). CHIPseq . Read more (PubMed: 28054639) »
  • Wang G  et al. Systematic analysis of the lysine acetylome reveals diverse functions of lysine acetylation in the oleaginous yeast Yarrowia lipolytica. AMB Express 7:94 (2017). WB . Read more (PubMed: 28497289) »

See all 59 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

Submitted Dec 12 2011

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

Sebastian Alvarado

Verified customer

Submitted 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

Submitted 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

Submitted 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

Submitted 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

Submitted 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

Submitted 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

Submitted 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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Thank you for your enquiry.

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.

One of my colleagues has previously compared the immunogen sequ...

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