Anti-Histone H3 抗体 [mAbcam 24834] - Nuclear Loading Control and ChIP Grade (ab24834)

製品の概要

  • 製品名
    Anti-Histone H3 antibody [mAbcam 24834] - Nuclear Loading Control and ChIP Grade
    Histone H3 一次抗体 製品一覧
  • 製品の詳細
    Mouse monoclonal [mAbcam 24834] to Histone H3 - Nuclear Loading Control and ChIP Grade
  • アプリケーション
    適用あり: ChIP, WBmore details
  • 種交差性
    交差種: Mouse, Rat, Human
    交差が予測される動物種: a wide range of other species
  • 免疫原

    Synthetic peptide corresponding to Human Histone H3 aa 100 to the C-terminus.
    Database link: P68431
    (Peptide available as ab12149)

  • ポジティブ・コントロール
    • This antibody gave a positive signal in the following lysates: HeLa Histone Preparation; NIH3T3 whole cell; PC12 whole cell; Mouse Testis Tissue; Rat Testis Tissue. This antibody gave a positive result in IHC in the following FFPE tissue: Human normal skin.
  • 特記事項

    This antibody clone is manufactured by Abcam.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab24834 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
ChIP Use 5 µg for 25 µg of chromatin.
WB Use a concentration of 0.5 - 1 µg/ml. Detects a band of approximately 18 kDa (predicted molecular weight: 15 kDa).Can be blocked with Human Histone H3 peptide (ab12149).

ターゲット情報

  • 機能
    Core component of nucleosome. 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.
  • 配列類似性
    Belongs to the histone H3 family.
  • 発生段階
    Expressed during S phase, then expression strongly decreases as cell division slows down during the process of differentiation.
  • 翻訳後修飾
    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.
    Methylation at Lys-5 (H3K4me), Lys-37 (H3K36me) and Lys-80 (H3K79me) are linked to gene activation. 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) are linked to gene repression. 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.
    Monoubiquitinated by RAG1 in lymphoid cells, monoubiquitination is required for V(D)J recombination (By similarity). Ubiquitinated by the CUL4-DDB-RBX1 complex in response to ultraviolet irradiation. This may weaken the interaction between histones and DNA and facilitate DNA accessibility to repair proteins.
  • 細胞内局在
    Nucleus. Chromosome.
  • Information by UniProt
  • 参照データベース
  • 別名
    • H3 histone family, member A antibody
    • H3/A antibody
    • H31_HUMAN antibody
    • H3FA antibody
    • Hist1h3a antibody
    • HIST1H3B antibody
    • HIST1H3C antibody
    • HIST1H3D antibody
    • HIST1H3E antibody
    • HIST1H3F antibody
    • HIST1H3G antibody
    • HIST1H3H antibody
    • HIST1H3I antibody
    • HIST1H3J antibody
    • histone 1, H3a antibody
    • Histone cluster 1, H3a antibody
    • Histone H3.1 antibody
    • Histone H3/a antibody
    • Histone H3/b antibody
    • Histone H3/c antibody
    • Histone H3/d antibody
    • Histone H3/f antibody
    • Histone H3/h antibody
    • Histone H3/i antibody
    • Histone H3/j antibody
    • Histone H3/k antibody
    • Histone H3/l antibody
    see all

画像

  • Chromatin was prepared from HeLa cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 5µg of ab24834 (blue), and 20µl 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.

  • Anti-Histone H3 antibody [mAbcam 24834] - Nuclear Loading Control and ChIP Grade (ab24834) at 1 µg/ml + Hela histone prep at 1 µg/ml

    Secondary
    Rabbit Anti-Mouse IgG H&L (HRP) (ab6728) at 1/5000 dilution

    Predicted band size : 15 kDa
    Observed band size : 18 kDa (why is the actual band size different from the predicted?)
  • All lanes : Anti-Histone H3 antibody [mAbcam 24834] - Nuclear Loading Control and ChIP Grade (ab24834) at 1 µg/ml

    Lane 1 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lane 2 : Testis (Mouse) Tissue Lysate
    Lane 3 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate
    Lane 4 : Testis (Rat) Tissue Lysate - normal tissue (ab29388)

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 15 kDa
    Observed band size : 18 kDa (why is the actual band size different from the predicted?)


    Exposure time : 3 minutes

プロトコール

参考文献

This product has been referenced in:
  • Anderson L  et al. Histone deacetylase inhibition modulates histone acetylation at gene promoter regions and affects genome-wide gene transcription in Schistosoma mansoni. PLoS Negl Trop Dis 11:e0005539 (2017). WB, ChIP ; Schistosoma mansoni . Read more (PubMed: 28406899) »
  • Bressan RB  et al. Efficient CRISPR/Cas9-assisted gene targeting enables rapid and precise genetic manipulation of mammalian neural stem cells. Development 144:635-648 (2017). WB ; Human . Read more (PubMed: 28096221) »

See all 11 Publications for this product

レビューと Q&A

Application
Western blot
Sample
Mouse Cell lysate - whole cell (primary cortical neurons)
Gel Running Conditions
Reduced Denaturing (4-12%Bis-Tris)
Loading amount
10 µg
Specification
primary cortical neurons
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

投稿 Aug 17 2016

Application
Western blot
Sample
Saccharomyces cerevisiae Cell lysate - nuclear (Histones)
Gel Running Conditions
Reduced Denaturing (12)
Loading amount
100 µg
Specification
Histones
Blocking step
Odyssey Blocking buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 50% · Temperature: 25°C
Username

David Truong

Verified customer

投稿 Sep 10 2015

Application
Western blot
Sample
Schizosaccharomyces pombe Cell lysate - whole cell (S. pombe and S. cerevisiae)
Gel Running Conditions
Reduced Denaturing (15%)
Loading amount
1e+007 cells
Specification
S. pombe and S. cerevisiae
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

投稿 Sep 09 2015

Application
Western blot
Loading amount
3e+007 cells
Gel Running Conditions
Non-reduced Denaturing (20% SDS-PAGE)
Sample
Plasmodium falciparum Cell lysate - whole cell (intraerythrocitic stages, synchronized culture)
Specification
intraerythrocitic stages, synchronized culture
Blocking step
Pierce Protein-Free T20 Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: RT°C
Username

Abcam user community

Verified customer

投稿 Mar 30 2015

Application
Western blot
Loading amount
10 µg
Gel Running Conditions
Reduced Denaturing (10 %)
Sample
Rat Cell lysate - nuclear (Brain-striatum and cortex)
Specification
Brain-striatum and cortex
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Mr. Tianyu Zhang

Verified customer

投稿 Jan 21 2015

All of our products are for research purposes only and are not certified for clinical research applications.

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (HEK293)
Loading amount
5 µg
Specification
HEK293
Treatment
50nM TSA for 24hrs
Gel Running Conditions
Non-reduced Denaturing (4-12%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

投稿 Jul 22 2010

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

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