Anti-Histone H3.3抗体- ChIP Grade (ab62642)

概述

  • 产品名称Anti-Histone H3.3抗体- ChIP Grade
    参阅全部 Histone H3.3 一抗
  • 描述
    兔多克隆抗体to Histone H3.3 - ChIP Grade
  • 经测试应用适用于: ELISA, ICC/IF, IHC-P, WB, ChIPmore details
  • 种属反应性
    与反应: Rat, Human
  • 免疫原

    Synthetic peptide corresponding to Human Histone H3.3.
    Database link: P84243

  • 阳性对照
    • Hela cells and human brain tissue

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. Avoid freeze / thaw cycle.
  • 存储溶液Preservative: 0.02% Sodium Azide
    Constituents: 50% Glycerol, PBS, 150mM Sodium chloride, pH 7.4
  • Concentration information loading...
  • 纯度Immunogen affinity purified
  • 纯化说明The antibody was affinity purified from rabbit antiserum by affinity chromatography using epitope specific immunogen.
  • 克隆多克隆
  • 同种型IgG
  • 研究领域

应用

Our Abpromise guarantee covers the use of ab62642 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
ELISA 1/5000.
ICC/IF 1/500 - 1/1000.
IHC-P 1/50 - 1/100.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 19 kDa (predicted molecular weight: 15 kDa).
ChIP Use at an assay dependent concentration.

靶标

  • 功能Variant histone H3 which replaces conventional H3 in a wide range of nucleosomes in active genes. Constitutes the predominant form of histone H3 in non-dividing cells and is incorporated into chromatin independently of DNA synthesis. Deposited at sites of nucleosomal displacement throughout transcribed genes, suggesting that it represents an epigenetic imprint of transcriptionally active chromatin. 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.
  • 序列相似性Belongs to the histone H3 family.
  • 发展阶段Expressed throughout the cell cycle independently of DNA synthesis.
  • 翻译后修饰Acetylation is generally linked to gene activation. Acetylation on Lys-10 (H3K9ac) impairs methylation at Arg-9 (H3R8me2s). Acetylation on Lys-19 (H3K18ac) and Lys-24 (H3K24ac) favors methylation at Arg-18 (H3R17me).
    Citrullination at Arg-9 (H3R8ci) and/or Arg-18 (H3R17ci) by PADI4 impairs methylation and represses transcription.
    Asymmetric dimethylation at Arg-18 (H3R17me2a) by CARM1 is linked to gene activation. Symmetric dimethylation at Arg-9 (H3R8me2s) by PRMT5 is linked to gene repression. Asymmetric dimethylation at Arg-3 (H3R2me2a) by PRMT6 is linked to gene repression and is mutually exclusive with H3 Lys-5 methylation (H3K4me2 and H3K4me3). H3R2me2a is present at the 3' of genes regardless of their transcription state and is enriched on inactive promoters, while it is absent on active promoters.
    Specifically enriched in modifications associated with active chromatin such as methylation at Lys-5 (H3K4me), Lys-37 and Lys-80. Methylation at Lys-5 (H3K4me) facilitates subsequent acetylation of H3 and H4. Methylation at Lys-80 (H3K79me) is associated with DNA double-strand break (DSB) responses and is a specific target for TP53BP1. Methylation at Lys-10 (H3K9me) and Lys-28 (H3K27me), which are linked to gene repression, are underrepresented. Methylation at Lys-10 (H3K9me) is a specific target for HP1 proteins (CBX1, CBX3 and CBX5) and prevents subsequent phosphorylation at Ser-11 (H3S10ph) and acetylation of H3 and H4. Methylation at Lys-5 (H3K4me) and Lys-80 (H3K79me) require preliminary monoubiquitination of H2B at 'Lys-120'. Methylation at Lys-10 (H3K9me) and Lys-28 (H3K27me) are enriched in inactive X chromosome chromatin.
    Phosphorylated at Thr-4 (H3T3ph) by GSG2/haspin during prophase and dephosphorylated during anaphase. Phosphorylation at Ser-11 (H3S10ph) by AURKB is crucial for chromosome condensation and cell-cycle progression during mitosis and meiosis. In addition phosphorylation at Ser-11 (H3S10ph) by RPS6KA4 and RPS6KA5 is important during interphase because it enables the transcription of genes following external stimulation, like mitogens, stress, growth factors or UV irradiation and result in the activation of genes, such as c-fos and c-jun. Phosphorylation at Ser-11 (H3S10ph), which is linked to gene activation, prevents methylation at Lys-10 (H3K9me) but facilitates acetylation of H3 and H4. Phosphorylation at Ser-11 (H3S10ph) by AURKB mediates the dissociation of HP1 proteins (CBX1, CBX3 and CBX5) from heterochromatin. Phosphorylation at Ser-11 (H3S10ph) is also an essential regulatory mechanism for neoplastic cell transformation. Phosphorylated at Ser-29 (H3S28ph) by MLTK isoform 1, RPS6KA5 or AURKB during mitosis or upon ultraviolet B irradiation. Phosphorylation at Thr-7 (H3T6ph) by PRKCBB is a specific tag for epigenetic transcriptional activation that prevents demethylation of Lys-5 (H3K4me) by LSD1/KDM1A. At centromeres, specifically phosphorylated at Thr-12 (H3T11ph) from prophase to early anaphase, by DAPK3 and PKN1. Phosphorylation at Thr-12 (H3T11ph) by PKN1 is a specific tag for epigenetic transcriptional activation that promotes demethylation of Lys-10 (H3K9me) by KDM4C/JMJD2C. Phosphorylation at Tyr-42 (H3Y41ph) by JAK2 promotes exclusion of CBX5 (HP1 alpha) from chromatin. Phosphorylation on Ser-32 (H3S31ph) is specific to regions bordering centromeres in metaphase chromosomes.
    Ubiquitinated. Monoubiquitinated by RAG1 in lymphoid cells, monoubiquitination is required for V(D)J recombination.
  • 细胞定位Nucleus. Chromosome.
  • Information by UniProt
  • 数据库链接
  • 别名
    • H3 histone family 3A antibody
    • H3 histone family 3B antibody
    • H3 histone, family 3B (H3.3B) antibody
    • H3.3 antibody
    • H3.3A antibody
    • H3.3B antibody
    • H33_HUMAN antibody
    • H3F3 antibody
    • H3F3A antibody
    • H3f3b antibody
    • Histone H3.3 antibody
    • Histone H3.3Q antibody
    • Histone H3.A antibody
    • Histone H3.B antibody
    • MGC87782 antibody
    • MGC87783 antibody
    see all

Anti-Histone H3.3 antibody - ChIP Grade 图像

  • Immunofluorescence analysis of HeLa cells, using ab62642 at a 1/500 dilution.
    Left image untreated.
    Right image treated with peptide.
  • Immunohistochemistry analysis of paraffin embedded human brain tissue using ab62642 at a 1/50 dilution.
    Left image untreated.
    Right image treated with peptide.
  • ChIP of Histone H3.3 from primary adipose chromatin using ab62642, showing enrichment at promoter regions previously associated with Histone H3.3 enrichment. The ChIP experiment was performed according to Dahl and Collas, 2007 (please see accompanying abreview for further details).

    See Abreview

  • Anti-Histone H3.3 antibody - ChIP Grade (ab62642) at 1 µg/ml + A431 (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 15 kDa
    Observed band size : 19 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 45 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 12 minutes

Anti-Histone H3.3 antibody - ChIP Grade (ab62642)参考文献

This product has been referenced in:
  • Su CH  et al. An H2A Histone Isotype, H2ac, Associates with Telomere and Maintains Telomere Integrity. PLoS One 11:e0156378 (2016). ChIP ; Human . Read more (PubMed: 27228173) »
  • Lee B  et al. Synergistic activation of Arg1 gene by retinoic acid and IL-4 involves chromatin remodeling for transcription initiation and elongation coupling. Nucleic Acids Res 44:7568-79 (2016). Read more (PubMed: 27166374) »

See all 11 Publications for this product

Product Wall

Abcam has not validated the combination of species/application used in this Abreview.
Application Western blot
Loading amount 20000 cells
Gel Running Conditions Reduced Denaturing (NuPAGE Bis-Tris gel 4-12%)
Sample Zebrafish Cell lysate - whole cell (whole embryo, deyolked)
Specification whole embryo, deyolked
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Abcam user community

Verified customer

提交于 Jun 24 2013

Abcam has not validated the combination of species/application used in this Abreview.
Application Western blot
Sample Mouse Cell lysate - whole cell (Raw 264.7)
Loading amount 25 µg
Specification Raw 264.7
Treatment DRB and LPS 0-2hrs
Gel Running Conditions Reduced Denaturing (12)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
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Abcam user community

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提交于 Jul 31 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (osteosarcoma cell line, Saos2)
Specification osteosarcoma cell line, Saos2
Fixative Paraformaldehyde
Permeabilization Yes - PBS TRITON .25%
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
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Abcam user community

Verified customer

提交于 Apr 18 2012

To our knowledge, ab62642 has not been tested in bull. Therefore, I can offer a discount off a future purchase if you buy ab62642 now, test it inbull and submit feedback to us in the form of an Abreview. It doesn't matter whether the Abreview is positi...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application ChIP
Sample Human Cell lysate - nuclear (Colon cancer cell)
Specification Colon cancer cell
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Detection step Real-time PCR
Negative control IgG control
Username

Dr. santosh chauhan

Verified customer

提交于 Aug 02 2011

Abcam has not validated the combination of species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Zebra finch Cell (HEK293 transfected with zebra finch H3.3 (GFP))
Specification HEK293 transfected with zebra finch H3.3 (GFP)
Fixative Paraformaldehyde
Permeabilization Yes - 0.3% TritonX-100
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 May 27 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application ChIP
Sample Human Cell lysate - whole cell (Soluble chromatin preparation from primary adipose)
Specification Soluble chromatin preparation from primary adipose
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 8 minute(s) and 0 second(s)
Specification of the cross-linking agent: Formaldehyde, 1%
Detection step Real-time PCR
Positive control Promoter regions for which we have strong evidence of epitope tagged H3.3 enrichment in the human genome data provided byh ChIP-on-chip and ChIP-qPCR.
Negative control No antibody Dynabeads Protein A-only control
Username

Prof. Philippe Collas

Verified customer

提交于 Oct 01 2009

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"