製品の概要

  • 製品名
    Anti-Histone H3 (tri methyl K79) antibody - ChIP Grade
    Histone H3 一次抗体 製品一覧
  • 製品の詳細
    Rabbit polyclonal to Histone H3 (tri methyl K79) - ChIP Grade
  • 由来種
    Rabbit
  • 特異性
    Reacts with Tri-methyl K79 of histone H3. Cross-reactivity to di-methyl K79.
  • アプリケーション
    適用あり: ICC/IF, ChIP, WB, ChIP/Chipmore details
  • 種交差性
    交差種: Mouse, Rat, Cow, Human, Saccharomyces cerevisiae, Plasmodium falciparum
  • 免疫原

    Synthetic peptide within Human Histone H3 aa 50 to the C-terminus (tri methyl K79) conjugated to Keyhole Limpet Haemocyanin (KLH). The exact sequence is proprietary.
    (Peptide available as ab4557)

  • ポジティブ・コントロール
    • WB: Calf thymus histone preparation and HeLa whole cell extract.

製品の特性

  • 製品の状態
    Liquid
  • 保存方法
    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.
  • バッファー
    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...
  • 精製度
    Immunogen affinity purified
  • ポリ/モノ
    ポリクローナル
  • アイソタイプ
    IgG
  • 研究分野

アプリケーション

Our Abpromise guarantee covers the use of ab2621 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
ICC/IF Use at an assay dependent concentration.
ChIP Use 2-4 µg for 6 µg of chromatin.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa).Can be blocked with Human Histone H3 (tri methyl K79) peptide (ab4557).

Abcam recommends using BSA as the blocking agent.

ChIP/Chip Use at an assay dependent concentration.

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 E pseudogene antibody
    • H3 histone family, member A antibody
    • H3/A antibody
    • H31_HUMAN antibody
    • H3F3 antibody
    • H3FA antibody
    • Hist1h3a antibody
    • HIST1H3B antibody
    • HIST1H3C antibody
    • HIST1H3D antibody
    • HIST1H3E antibody
    • HIST1H3F antibody
    • HIST1H3G antibody
    • HIST1H3H antibody
    • HIST1H3I antibody
    • HIST1H3J antibody
    • HIST3H3 antibody
    • histone 1, H3a antibody
    • Histone cluster 1, H3a antibody
    • Histone H3 3 pseudogene 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 U2OS 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  ab2621 (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. Primers and probes are located in the first kb of the transcribed region.

  • Immunocytochemistry/ Immunofluorescence analysis of HEK293 cells labeling Histone H3 (tri methyl K79) with ab2621 at 1/1000 dilution. Cells were fixed with paraformaldehyde and permeabilized with Triton-X. The cells were blocked with 5% BSA for 1 hour at 21°C, followed by incubation with Anti-Histone H3 (tri methyl K79) antibody - ChIP Grade (ab2621) in 3% BSA in TBST for 16 hours at 4°C. A goat anti-rabbit Alexa Fluro 594 secondary antibody was used at 1/1000 dilution.

    See Abreview

  • All lanes : Anti-Histone H3 (tri methyl K79) antibody - ChIP Grade (ab2621) at 1 µg/ml

    Lane 1 : Calf Thymus Histone Preparation Nuclear Lysate
    Lane 2 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (unmodified ) peptide (ab4558) at 0.5 µg/ml
    Lane 3 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (mono methyl K79) peptide (ab4555) at 0.5 µg/ml
    Lane 4 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (di methyl K79) peptide (ab4556) at 0.5 µg/ml
    Lane 5 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (tri methyl K79) peptide (ab4557) at 0.5 µg/ml
    Lane 6 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H4 (di methyl K79) peptide (ab4560) at 0.5 µg/ml
    Lane 7 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (di methyl K9) peptide (ab1772) at 0.5 µg/ml

    Lysates/proteins at 0.5 µg per lane.

    Secondary
    All lanes : IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size: 15 kDa
    Observed band size: 17 kDa
    why is the actual band size different from the predicted?

  • ab2621 (1/400) staining Histone H3 (tri Methyl K79) in human hTERT cells (green). Cells were fixed in methanol, permeabilised with 0.5% Triton X100/PBS and counterstained with DAPI in order to highlight the nucleus (red). For further experimental details, or to check results obtained using paraformaldehyde fixation, please refer to abreview.

    See Abreview

  • Chromatin was prepared from whole cell lysate of normal rat liver and liver cancer cells. The cross-linking (X-ChiP) technique was used, crosslinking was performed for 15 minutes in formaldehyde. 5 µg of the primary antibody was used in 1/100 dilution and it was incubated with the sample for 16 hours at 4°C. The immunoprecipitated DNA was quantified by real time PCR.

    See Abreview

参考文献

This product has been referenced in:
See all 102 Publications for this product

レビューと Q&A

1-10 of 15 Abreviews or Q&A

Application
Western blot
Sample
Fruit fly (Drosophila melanogaster) Tissue lysate - nuclear (whole fruit fly)
Gel Running Conditions
Reduced Denaturing
Loading amount
4.5 µg
Specification
whole fruit fly
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Mr. Alexander Hambly

Verified customer

投稿 Apr 14 2015

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Sample
Human Cell (HEK293)
Specification
HEK293
Permeabilization
Yes - TRITON
Fixative
Paraformaldehyde

Abcam user community

Verified customer

投稿 Jun 20 2014

Application
ChIP
Detection step
Real-time PCR
Sample
Human Cell lysate - whole cell (SCC9 and OKF6)
Specification
SCC9 and OKF6
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

Mr. Dan Stummer

Verified customer

投稿 Apr 11 2014

Answer

Here is the WB and ChIP data for lot GR65540-1 (attached) as well as the reviewers notes below. The WB image and ChIP data are consistent with that shown on the datasheet. The only difference is the low signal with ALDOA in ChIP but the stock control run at the same time showed the same low signal so the reviewer wasn’t concerned.
7. WB pass. Lot pass. Pass WB - good blocking with immunising peptide. Can see xr with diK79 peptide but datasheet mentions so allowed. Waiting on ChIP results. SL 22.12.09 Passed in ChIP. The GAPDH signal is variable in these HeLa's and the ALDOA signal is extremely low (even in the STOCK) so this loci is to be ignored in analysis. Passing the ChIP as the test AP is stronger than the STOCK and there is a 2 fold difference between the active and inactive enrichment. DRC 10/01/2010.
WB image caption:



All lanes : Anti-Histone H3 (tri methyl K79) antibody - ChIP Grade (ab2621) at 1 µg/ml

Lane 1 : Calf Thymus Histone Preparation Nuclear Lysate (ab121)
Lane 2 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with http://lab.adminsite.com/index.html?datasheet=4558 at 0.5 µg/ml
Lane 3 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with http://lab.adminsite.com/index.html?datasheet=1772 at 0.5 µg/ml
Lane 4 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with http://lab.adminsite.com/index.html?datasheet=4555 at 0.5 µg/ml
Lane 5 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with http://lab.adminsite.com/index.html?datasheet=4556 at 0.5 µg/ml
Lane 6 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with http://lab.adminsite.com/index.html?datasheet=4557 at 0.5 µg/ml
Lane 7 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with http://lab.adminsite.com/index.html?datasheet=4560 at 0.5 µg/ml

Lysates/proteins at 0.5 µg per lane.

Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) (ab65484) at 1/3000 dilution

Performed under reducing conditions.

Predicted band size : 15 kDa


Exposure time : 90 seconds

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
ChIP
Sample
Human Cell lysate - nuclear (HEK293)
Specification
HEK293
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 20 minute(s) and 0 second(s)
Specification of the cross-linking agent: 1% Formaldehyde
Detection step
Real-time PCR
Negative control
No antibody

Abcam user community

Verified customer

投稿 Jan 30 2012

Answer

We are not able to "save" specific vials of antibody for customer that may wish to purchase them in the future. If you'd like I can recommend specific lots that have the most stock, which may give you the best chance to buy an antibody at a later date from that same stock. The downside with this scenario is that it will only increase the likelihood the specific lot will be available but not guarantee it.

Also, perhaps shortening the time frame for testing the antibodies might also give us a better chance to provide you with the lots you need.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - whole cell (Mouse ES cells)
Loading amount
30 µg
Specification
Mouse ES cells
Gel Running Conditions
Reduced Denaturing (15% SDS-PAGE)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: RT°C

Abcam user community

Verified customer

投稿 Nov 07 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (Hela Cells)
Loading amount
50 µg
Specification
Hela Cells
Gel Running Conditions
Reduced Denaturing (15% SDS-PAGE)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: RT°C

Abcam user community

Verified customer

投稿 Nov 07 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
ChIP
Sample
Mouse Cell lysate - whole cell (teratocarcinoma F9 cells)
Specification
teratocarcinoma F9 cells
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 0 second(s)
Specification of the cross-linking agent: DSG and formaldehyde
Detection step
Real-time PCR
Positive control
H3K4 trimethyl
Negative control
Rabbit IgG

Mr. Dan Stummer

Verified customer

投稿 Mar 15 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - nuclear (Liver)
Loading amount
20 µg
Specification
Liver
Treatment
X protein was knockdowned
Gel Running Conditions
Reduced Denaturing (15)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

投稿 Feb 24 2011

1-10 of 15 Abreviews or Q&A

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

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