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    histone-h2b-mono-methyl-k5-antibody-chip-grade-ab12929.pdf

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Epigenetics and Nuclear Signaling Histones H2B Methylated
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Anti-Histone H2B (mono methyl K5) 抗体 - ChIP Grade (ab12929)

  • Datasheet
Reviews (5)Q&A (1)References (4)

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ChIP - Anti-Histone H2B (mono methyl K5) antibody - ChIP Grade (ab12929)
  • Western blot - Anti-Histone H2B (mono methyl K5) antibody - ChIP Grade (ab12929)
  • ChIP - Anti-Histone H2B (mono methyl K5) antibody - ChIP Grade (ab12929)
  • Immunoprecipitation - Anti-Histone H2B (mono methyl K5) antibody - ChIP Grade (ab12929)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2B (mono methyl K5) antibody - ChIP Grade (ab12929)

Key features and details

  • Rabbit polyclonal to Histone H2B (mono methyl K5) - ChIP Grade
  • Suitable for: IHC-P, IP, WB, ChIP
  • Reacts with: Cow, Human
  • Isotype: IgG

こちらの製品もご検討ください

二次抗体
Product image
Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
一次抗体
Product image
Anti-TNP1 antibody (ab73135)
タンパク質
Product image
Recombinant Human Histone H2B protein (ab92409)

関連製品

製品の概要

  • 製品名

    Anti-Histone H2B (mono methyl K5) antibody - ChIP Grade
    Histone H2B 一次抗体 製品一覧
  • 製品の詳細

    Rabbit polyclonal to Histone H2B (mono methyl K5) - ChIP Grade
  • 由来種

    Rabbit
  • 特異性

    Mono methylation of Histone H2B K5 is a putative modification site. ab22512 in ELISA specifically recognises mono-methyl K5 histone H2B peptide but not the corresponding unmodified histone H2B peptide.
  • アプリケーション

    適用あり: IHC-P, IP, WB, ChIPmore details
  • 種交差性

    交差種: Cow, Human
    交差が予測される動物種: Mouse, Rat, Chicken, Xenopus laevis
  • 免疫原

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • 特記事項

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

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

    • Epigenetics and Nuclear Signaling
    • Histones
    • H2B
    • Methylated
    • Epigenetics and Nuclear Signaling
    • ChIP assays
    • ChIP antibodies

関連製品

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Recombinant Protein

    • Recombinant Human Histone H2B protein (ab92409)

アプリケーション

The Abpromise guarantee

Abpromise保証は、 次のテスト済みアプリケーションにおけるab12929の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション Abreviews 特記事項
IHC-P
Use a concentration of 0.1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IP
Use at an assay dependent concentration.
WB (3)
Use a concentration of 1 µg/ml.
ChIP (1)
Use 2-5 µg for 25 µg of chromatin.
特記事項
IHC-P
Use a concentration of 0.1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IP
Use at an assay dependent concentration.
WB
Use a concentration of 1 µg/ml.
ChIP
Use 2-5 µg for 25 µg of chromatin.

ターゲット情報

  • 関連性

    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. Subunit structure The nucleosome is a histone octamer containing two molecules each of H2A, H2B, H3 and H4 assembled in one H3-H4 heterotetramer and two H2A-H2B heterodimers. The octamer wraps approximately 147 bp of DNA. Post-translational modification Monoubiquitination at Lys-35 (H2BK34Ub) by the MSL1/MSL2 dimer is required for histone H3 'Lys-4' (H3K4me) and 'Lys-79' (H3K79me) methylation and transcription activation at specific gene loci, such as HOXA9 and MEIS1 loci. Similarly, monoubiquitination at Lys-121 (H2BK120Ub) by the RNF20/40 complex gives a specific tag for epigenetic transcriptional activation and is also prerequisite for histone H3 'Lys-4' and 'Lys-79' methylation. It also functions cooperatively with the FACT dimer to stimulate elongation by RNA polymerase II. H2BK120Ub also acts as a regulator of mRNA splicing: deubiquitination by USP49 is required for efficient cotranscriptional splicing of a large set of exons. Phosphorylation at Ser-37 (H2BS36ph) by AMPK in response to stress promotes transcription. Phosphorylated on Ser-15 (H2BS14ph) by STK4/MST1 during apoptosis; which facilitates apoptotic chromatin condensation. Also phosphorylated on Ser-15 in response to DNA double strand breaks (DSBs), and in correlation with somatic hypermutation and immunoglobulin class-switch recombination. GlcNAcylation at Ser-113 promotes monoubiquitination of Lys-121. It fluctuates in response to extracellular glucose, and associates with transcribed genes. Crotonylation (Kcr) is specifically present in male germ cells and marks testis-specific genes in post-meiotic cells, including X-linked genes that escape sex chromosome inactivation in haploid cells. Crotonylation marks active promoters and enhancers and confers resistance to transcriptional repressors. It is also associated with post-meiotically activated genes on autosomes.
  • 細胞内局在

    Nuclear
  • 参照データベース

    • Entrez Gene: 337874 Human
    • Entrez Gene: 54145 Human
    • Entrez Gene: 8349 Human
    • Entrez Gene: 104021 Mouse
    • Entrez Gene: 64647 Rat
    • SwissProt: P0C1H5 Chicken
    • SwissProt: P62808 Cow
    • SwissProt: O60814 Human
    • SwissProt: P06899 Human
    • SwissProt: P23527 Human
    • SwissProt: P33778 Human
    • SwissProt: P57053 Human
    • SwissProt: P58876 Human
    • SwissProt: P62807 Human
    • SwissProt: Q16778 Human
    • SwissProt: Q5QNW6 Human
    • SwissProt: Q8N257 Human
    • SwissProt: Q93079 Human
    • SwissProt: Q99877 Human
    • SwissProt: Q99879 Human
    • SwissProt: Q99880 Human
    • SwissProt: P10854 Mouse
    • SwissProt: Q64475 Mouse
    • SwissProt: Q00729 Rat
    see all
  • 別名

    • GL105 antibody
    • H2B GL105 antibody
    • H2B histone family member O antibody
    • H2B histone family member S antibody
    • H2B.1 antibody
    • H2B.1 B antibody
    • H2B.b antibody
    • H2B.c antibody
    • H2B.d antibody
    • H2B.e antibody
    • H2B.f antibody
    • H2B.j antibody
    • H2B.q antibody
    • H2B/b antibody
    • H2B/c antibody
    • H2B/d antibody
    • H2B/e antibody
    • H2B/f antibody
    • H2B/j antibody
    • H2B/o antibody
    • H2B/q antibody
    • H2BFB antibody
    • H2BFC antibody
    • H2BFD antibody
    • H2BFE antibody
    • H2BFF antibody
    • H2BFJ antibody
    • H2BFO antibody
    • H2BFQ antibody
    • H2BFS antibody
    • H2BGL105 antibody
    • H2BQ antibody
    • HIRIP2 antibody
    • HIST1H2BB antibody
    • HIST1H2BD antibody
    • HIST1H2BH antibody
    • HIST1H2BL antibody
    • HIST1H2BM antibody
    • HIST1H2BN antibody
    • HIST2H2BE antibody
    • histone cluster 2 antibody
    • Histone H2B antibody
    • histone H2B GL105 antibody
    • Histone H2B type 1 B antibody
    • Histone H2B type 1 D antibody
    • Histone H2B type 1 H antibody
    • Histone H2B type 1 L antibody
    • Histone H2B type 1 M antibody
    • Histone H2B type 1 N antibody
    • Histone H2B type 2 E antibody
    • Histone H2B.q antibody
    • histone protein antibody
    see all

画像

  • ChIP - Anti-Histone H2B (mono methyl K5) antibody - ChIP Grade (ab12929)
    ChIP - Anti-Histone H2B (mono methyl K5) antibody - ChIP Grade (ab12929)

    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 ab12929 (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). Primers and probes are located in the first kb of the transcribed region.

  • Western blot - Anti-Histone H2B (mono methyl K5) antibody - ChIP Grade (ab12929)
    Western blot - Anti-Histone H2B (mono methyl K5) antibody - ChIP Grade (ab12929)
    All lanes : Anti-Histone H2B (mono methyl K5) antibody - ChIP Grade (ab12929) at 1 µg/ml

    Lane 1 : Calf Thymus Histone Preparation Nuclear Lysate (ab121)
    Lane 2 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with Human Histone H2B (mono methyl K5) peptide (ab13211)
    Lane 3 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with Human Histone H2B peptide (ab13212)

    Lysates/proteins at 20 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: 14 kDa

  • ChIP - Anti-Histone H2B (mono methyl K5) antibody - ChIP Grade (ab12929)
    ChIP - Anti-Histone H2B (mono methyl K5) antibody - ChIP Grade (ab12929)This image is courtesy of an anonymous abreview.
    ab12929 at a 1/600 dilution for ChIP analysis of mouse dorsal skin epidermis whole tissue lysate, incubated for 15 hours at 4°C with ChIP dilution buffer. Cross-linking (X-ChIP) using 1% formaldehyde for 10 minutes.
    Detection step: Semiquantitative PCR.
    Negative control: Rabbit IgG.
    Cells untreated.

    See Abreview

  • Immunoprecipitation - Anti-Histone H2B (mono methyl K5) antibody - ChIP Grade (ab12929)
    Immunoprecipitation - Anti-Histone H2B (mono methyl K5) antibody - ChIP Grade (ab12929)

    Histone H2B (mono methyl K5) was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to Histone H2B (mono methyl K5) - ChIP Grade and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab12929.
    Secondary: Clean blot (HRP conjugate) at 1/1000 dilution.
    Band: 17kDa: Histone H2B (mono methyl K5).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2B (mono methyl K5) antibody - ChIP Grade (ab12929)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2B (mono methyl K5) antibody - ChIP Grade (ab12929)

    IHC image of Histone H2B (mono methyl K5) staining in human pancreatic adenocarcinoma formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab12929, 0.1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

プロトコール

  • PCR primers used to test ab12929 in ChIP at Abcam

Click here to view the general protocols

データシートおよび資料

  • SDS download

  • Datasheet download

    Download

参考文献 (4)

ab12929 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab12929 は 4 報の論文で使用されています。

  • Majocchi S  et al. Epigenetic regulatory elements associate with specific histone modifications to prevent silencing of telomeric genes. Nucleic Acids Res N/A:N/A (2013). ChIP . PubMed: 24071586
  • Santoro SW & Dulac C The activity-dependent histone variant H2BE modulates the life span of olfactory neurons. elife 1:e00070 (2012). ICC/IF ; Mouse . PubMed: 23240083
  • Wang Z  et al. Combinatorial patterns of histone acetylations and methylations in the human genome. Nat Genet 40:897-903 (2008). WB, ChIP . PubMed: 18552846
  • Barski A  et al. High-resolution profiling of histone methylations in the human genome. Cell 129:823-37 (2007). PubMed: 17512414

レビューと Q&A

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レビューを送る 質問を送る

1-6 of 6 Abreviews or Q&A

Western blot abreview for Anti-Histone H2B (mono methyl K5) antibody - ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing (15%)
Sample
Fruit fly (Drosophila melanogaster) Tissue lysate - nuclear (Fruit fly embryo (16hr))
Specification
Fruit fly embryo (16hr)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Read More

Abcam user community

Verified customer

投稿 Feb 09 2015

Western blot abreview for Anti-Histone H2B (mono methyl K5) antibody - ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing (12.5%)
Sample
Rat Cell lysate - nuclear (PC12 cell line)
Specification
PC12 cell line
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Read More

Abcam user community

Verified customer

投稿 Jan 08 2015

Immunocytochemistry abreview for Anti-Histone H2B (mono methyl K5) antibody - ChIP Grade

Good
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry
Blocking step
BSA as blocking agent for 20 minute(s) · Concentration: 3% · Temperature: 23°C
Sample
Human Cultured Cells (Human embryonic stem cells)
Specification
Human embryonic stem cells
Permeabilization
Yes - 0.3% Triton X-100
Fixative
Paraformaldehyde
Read More

Abcam user community

Verified customer

投稿 Jan 08 2014

Western blot abreview for Anti-Histone H2B (mono methyl K5) antibody - ChIP Grade

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing (15%)
Sample
Mouse Cell lysate - whole cell (Mouse embryonic fibroblasts)
Specification
Mouse embryonic fibroblasts
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Read More

Abcam user community

Verified customer

投稿 Jan 08 2014

ChIP abreview for Anti-Histone H2B (mono methyl K5) antibody - ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
Application
ChIP
Sample
Mouse Tissue lysate - whole (dorsal skin epidermis)
Specification
dorsal skin epidermis
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: 1% formaldehyde
Detection step
Semiquantitative PCR
Negative control
Rabbit IgG
Read More

Abcam user community

Verified customer

投稿 Feb 15 2010

Question

I am currently using antibodies purchased from your company to detect peptides via western blot.

I am using ab40886 and ab12929 to detect my peptides. I am using both antibodies at 1:1000 concentrations. My problem is with ab40886. It does not detect my peptide that contains the acetylated lysine 5 residue. Ab12929 on the other hand detects my peptide with mono-methylated lysine 5 very well. I

Read More

Abcam community

Verified customer

Asked on Jan 19 2012

Answer

Thank you for contacting Abcam.

I am sorry that you are having problems with ab40886 in western blotting.

I would advise that you increase your primary antibody concentration to 1/500 and incubate at 4C overnight.

The antibody is covered under our Abpromise for six months and is guaranteed to work in western blot. If we cannot resolve the issue you are having with the antibody then I would be happy to either send a replacement antibody or to process a refund.

Read More

Abcam Scientific Support

Answered on Jan 19 2012

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

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