Anti-Histone H3 (tri methyl K4)抗体[mAbcam1012] - ChIP Grade (ab1012)

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ChIP - Anti-Histone H3 (tri methyl K4) antibody [mAbcam1012] - ChIP Grade (ab1012)
  • Immunocytochemistry - Anti-Histone H3 (tri methyl K4) antibody [mAbcam1012] - ChIP Grade (ab1012)
  • ELISA - Anti-Histone H3 (tri methyl K4) antibody [mAbcam1012] - ChIP Grade (ab1012)
  • Peptide Array - Anti-Histone H3 (tri methyl K4) antibody [mAbcam1012] - ChIP Grade (ab1012)
  • Immunocytochemistry - Anti-Histone H3 (tri methyl K4) antibody [mAbcam1012] - ChIP Grade (ab1012)
  • Western blot - Anti-Histone H3 (tri methyl K4) antibody [mAbcam1012] - ChIP Grade (ab1012)
  • Flow Cytometry - Anti-Histone H3 (tri methyl K4) antibody [mAbcam1012] - ChIP Grade (ab1012)

Key features and details

  • Mouse monoclonal [mAbcam1012] to Histone H3 (tri methyl K4) - ChIP Grade
  • Suitable for: PepArr, ELISA, ChIP, Flow Cyt, WB, ICC
  • Reacts with: Rat, Cow, Human, Rice, Recombinant fragment
  • Isotype: IgG2b

概述

  • 产品名称

    Anti-Histone H3 (tri methyl K4)抗体[mAbcam1012] - ChIP Grade
    参阅全部 Histone H3 一抗

  • 描述

    小鼠单克隆抗体[mAbcam1012] to Histone H3 (tri methyl K4) - ChIP Grade

  • 宿主

    Mouse

  • 特异性

    By ELISA the antibody binds to the tri methyl K4 peptide and partially to di and mono methyl K4 peptides (there is <10% cross reactivity with di methyl K4). It does not bind to unmodified, mono, di or tri methyl K9 or di or tri methyl K27 peptides. Not suitable for blocking with milk in Western blot (see Application notes).

  • 经测试应用

    适用于: PepArr, ELISA, ChIP, Flow Cyt, WB, ICCmore details

  • 种属反应性

    与反应: Rat, Cow, Human, Rice, Recombinant fragment
    预测可用于: Mouse, Sheep, Xenopus laevis, Arabidopsis thaliana, Caenorhabditis elegans, Drosophila melanogaster, Schizosaccharomyces pombe, Zebrafish, Mammals

  • 免疫原

    Synthetic peptide corresponding to Human Histone H3 aa 1-100 (tri methyl K4) conjugated to keyhole limpet haemocyanin (Cysteine residue).
    (Peptide available as ab1342)

  • 阳性对照

    • ChIP: GAPDH primer pair ab267832. ICC: HeLa cells.

  • 常规说明

    Learn about ChIP assay kits, other ChIP antibodies, protocols and more in the ChIP assay guide.

    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

性能

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab1012于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
PepArr
Use a concentration of 0.5 - 2 µg/ml.
ELISA
Use at an assay dependent concentration.
ChIP (9)
Use 2-5 µg for 25 µg of chromatin.

Use GAPDH ChIP primer pair ab267832 as positive control.
Flow Cyt
Use 1µg for 106 cells.

ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.
WB (7)
Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa).Can be blocked with Human Histone H3 (tri methyl K4) peptide (ab1342). NOT SUITABLE for blocking with milk. Block in 5% BSA for 1 hour. Our labs have investigated the blocking conditions for this antibody and found that milk significantly decreases the signal and is therefore not a suitable blocking agent for this antibody (see image below).
ICC
Use a concentration of 5 µg/ml.
说明
PepArr
Use a concentration of 0.5 - 2 µg/ml.
ELISA
Use at an assay dependent concentration.
ChIP
Use 2-5 µg for 25 µg of chromatin.

Use GAPDH ChIP primer pair ab267832 as positive control.
Flow Cyt
Use 1µg for 106 cells.

ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.
WB
Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa).Can be blocked with Human Histone H3 (tri methyl K4) peptide (ab1342). NOT SUITABLE for blocking with milk. Block in 5% BSA for 1 hour. Our labs have investigated the blocking conditions for this antibody and found that milk significantly decreases the signal and is therefore not a suitable blocking agent for this antibody (see image below).
ICC
Use a concentration of 5 µg/ml.

靶标

  • 功能

    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.

  • 序列相似性

    Belongs to the histone H3 family.

  • 发展阶段

    Expressed during S phase, then expression strongly decreases as cell division slows down during the process of differentiation.

  • 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.
    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.

  • 细胞定位

    Nucleus. Chromosome.
  • Information by UniProt

  • 数据库链接

    see all

  • 别名

    • 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

图片

  • ChIP - Anti-Histone H3 (tri methyl K4) antibody [mAbcam1012] - ChIP Grade (ab1012)
    ChIP - Anti-Histone H3 (tri methyl K4) antibody [mAbcam1012] - ChIP Grade (ab1012)

    Chromatin was prepared from U2OS cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10min. The  ChIP was performed with 25 µg of chromatin, 5 µg of  ab1012 (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.

  • Immunocytochemistry - Anti-Histone H3 (tri methyl K4) antibody [mAbcam1012] - ChIP Grade (ab1012)
    Immunocytochemistry - Anti-Histone H3 (tri methyl K4) antibody [mAbcam1012] - ChIP Grade (ab1012)This image is courtesy of Darin McDonald, Cross Cancer Institute
    SK-N-SK cells were fixed in 4% paraformaldehdye, permeabilized in 0.5% Triton X-100 and incubated with ab1012 (1/100). The antibody clearly stains the nucleus (red). The cells were counterstained with DAPI (blue). 100x magnification.
  • ELISA - Anti-Histone H3 (tri methyl K4) antibody [mAbcam1012] - ChIP Grade (ab1012)
    ELISA - Anti-Histone H3 (tri methyl K4) antibody [mAbcam1012] - ChIP Grade (ab1012)

    ELISA using ab1012 at varying antibody concentrations.

    Curve_SPL1 indicates binding to the Human Histone H3 (tri methyl K4) peptide (ab1342). Curve_SPL4 indicates partial binding to the Human Histone H3 (di methyl K4) peptide (ab7768). There is very weak cross-reactivity with the Human Histone H3 (mono methyl K4) peptide (ab1340) (Curve_SPL3).

    Binding to the following peptides was not seen: SPL2 unmodified Histone H3, SPL5 Histone H3 mono methyl K9, SPL6 Histone H3 di methyl K9, SPL7 Histone H3 tri methyl K9, SPL8 Histone H3 mono methyl K27, SPL9 Histone H3 di methyl K27, SPL10 Histone H3 tri methyl K27.

  • Peptide Array - Anti-Histone H3 (tri methyl K4) antibody [mAbcam1012] - ChIP Grade (ab1012)
    Peptide Array - Anti-Histone H3 (tri methyl K4) antibody [mAbcam1012] - ChIP Grade (ab1012)

    All batches of ab1012 are tested in Peptide Array against peptides to different Histone modifications. Six dilutions of each peptide are printed on to the Peptide Array in triplicate and results are averaged before being plotted on to a graph. Results show strong binding to Histone H3 - tri methyl K4 peptide (ab92374), indicating that this antibody specifically recognises the Histone H3 - tri methyl K4 modification.

    ab92374 - Histone H3 - tri methyl K4
    ab7768 - Histone H3 - di methyl K4
    ab179150 - Histone H3 - asymmetric di methyl R2
    ab179142 - Histone H3 - mono methyl R2
    ab1784 - Histone H3 - di methyl K36
    ab179146 - Histone H3 - symmetric di methyl R2
    ab36927 - Histone H2A - phospho S122
    ab46854 - Histone H3 - tri methyl K18
    ab179021 - Histone H2A - symmetric di methyl R29
    ab17632 - Histone H4 - biotinylated K5
    ab179159 - Histone H3 - phospho T3
    ab14103 - Histone H3 - phopsho T6
    ab1783 - Histone H3 - mono methyl K36
    ab179177 - Histone H3 - mono methyl K4 + Histone H3 di methyl K9
  • Immunocytochemistry - Anti-Histone H3 (tri methyl K4) antibody [mAbcam1012] - ChIP Grade (ab1012)
    Immunocytochemistry - Anti-Histone H3 (tri methyl K4) antibody [mAbcam1012] - ChIP Grade (ab1012)

    ab1012 staining Histone H3 (Tri methyl K4) in HeLa cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab1012 at 5µg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

    Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

  • Western blot - Anti-Histone H3 (tri methyl K4) antibody [mAbcam1012] - ChIP Grade (ab1012)
    Western blot - Anti-Histone H3 (tri methyl K4) antibody [mAbcam1012] - ChIP Grade (ab1012)
    Lane 1 : Anti-Histone H3 (tri methyl K4) antibody [mAbcam1012] - ChIP Grade (ab1012) at 1 µg/ml (Blocked in 5% BSA)
    Lane 2 : Anti-Histone H3 (tri methyl K4) antibody [mAbcam1012] - ChIP Grade (ab1012) at 1 µg/ml (Blocked in 5% MILK)

    All lanes : Calf Thymus Histone Preparation Nuclear Lysate

    Lysates/proteins at 0.5 µg per lane.

    Secondary
    All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 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?


    Exposure time: 10 minutes
  • Flow Cytometry - Anti-Histone H3 (tri methyl K4) antibody [mAbcam1012] - ChIP Grade (ab1012)
    Flow Cytometry - Anti-Histone H3 (tri methyl K4) antibody [mAbcam1012] - ChIP Grade (ab1012)

    Overlay histogram showing HeLa cells stained with ab1012 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab1012, 1µg/1x106) for 30 min at 22°C.  The secondary antibody used was Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117) at 1/2000 dilution for 30 min at 22°C.

    Isotype control antibody (black line) was Mouse IgG2b [7E10G10] isotype control (ab170192) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

    Acquisition of >5,000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter.

    This antibody gave a positive signal in HeLa cells fixed with 4% formaldehyde (10 min)/permeabilized with 0.1% PBS-Triton X-100 for 15 min used under the same conditions.

文献 (167)

发表研究结果有使用 ab1012？请让我们知道，以便我们可以引用本数据表中的参考文章。

ab1012 被引用在 167 文献中.

  • Ghasemi R  et al. Contribution of CTCF binding to transcriptional activity at the HOXA locus in NPM1-mutant AML cells. Leukemia N/A:N/A (2020). PubMed: 32398790
  • Kim CY  et al. The LncRNA HOTAIRM1 Promotes Tamoxifen Resistance by Mediating HOXA1 Expression in ER+ Breast Cancer Cells. J Cancer 11:3416-3423 (2020). PubMed: 32284737
  • González B  et al. Dopamine Receptor D1 Contributes to Cocaine Epigenetic Reprogramming of Histone Modifications in Male Germ Cells. Front Cell Dev Biol 8:216 (2020). PubMed: 32318569
  • Zhang W  et al. SWI/SNF complex subunit BAF60a represses hepatic ureagenesis through a crosstalk between YB-1 and PGC-1a. Mol Metab 32:85-96 (2020). PubMed: 32029232
  • Li X  et al. Epigenetic Regulation of Key Enzymes CYP7a1 and HMGCR Affect Hepatic Cholesterol Metabolism in Different Breeds of Piglets. Front Vet Sci 7:231 (2020). PubMed: 32500085
View all Publications for this product

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1-7 of 7 Abreviews or Q&A

Western blot abreview for Anti-Histone H3 (tri methyl K4) antibody [mAbcam1012] - ChIP Grade

 Good
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (Mammary cell line)
Gel Running Conditions
Reduced Denaturing (10)
Loading amount
1e+006 cells
Specification
Mammary cell line
Blocking step
Milk as blocking agent for 40 minute(s) · Concentration: 5% · Temperature: 25°C
Read More

Abcam user community

Verified customer

提交于 Feb 28 2017

Western blot abreview for Anti-Histone H3 (tri methyl K4) antibody [mAbcam1012] - ChIP Grade

 Good
Abreviews
abreview image
Application
Western blot
Loading amount
1e+006 cells
Gel Running Conditions
Reduced Denaturing (12%)
Sample
Human Cell lysate - whole cell (Breast)
Specification
Breast
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Read More

Abcam user community

Verified customer

提交于 Sep 01 2014

Western blot abreview for Anti-Histone H3 (tri methyl K4) antibody [mAbcam1012] - ChIP Grade

 Good
Abreviews
abreview image
Application
Western blot
Loading amount
1e+006 cells
Gel Running Conditions
Reduced Denaturing (12%)
Sample
Mouse Cell lysate - whole cell (Colon)
Specification
Colon
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Read More

Abcam user community

Verified customer

提交于 Sep 01 2014

Western blot abreview for Anti-Histone H3 (methylated) antibody [mAbcam1012] - ChIP Grade

 Poor
Abreviews
abreview image
Application
Western blot
Sample
Saccharomyces cerevisiae Cell lysate - whole cell (lysate prepared from 1 OD of cells from liquid cul)
Loading amount
17 µg
Specification
lysate prepared from 1 OD of cells from liquid cul
Gel Running Conditions
Reduced Denaturing (12%)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: RT°C
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Dr. Marc Meneghini

Verified customer

提交于 Apr 20 2011

Western blot abreview for Anti-Histone H3 (methylated) antibody [mAbcam1012] - ChIP Grade

 Poor
Abreviews
abreview image
Application
Western blot
Sample
Saccharomyces cerevisiae Cell lysate - nuclear (yeast)
Loading amount
5 µg
Specification
yeast
Gel Running Conditions
Reduced Denaturing (16% gel)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More

Jessica Jackson

Verified customer

提交于 Nov 30 2010

Western blot abreview for Anti-Histone H3 (methylated) antibody [mAbcam1012] - ChIP Grade

 Inconclusive
Abreviews
abreview image
Application
Western blot
Sample
Mouse Tissue lysate - whole (E9.5 embryo)
Loading amount
20 µg
Specification
E9.5 embryo
Gel Running Conditions
Non-reduced Denaturing (16% acrylamide)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More

Dr. Matthieu Boulard

Verified customer

提交于 Aug 20 2010

Western blot abreview for Anti-Histone H3 (methylated) antibody [mAbcam1012] - ChIP Grade

 Average
Abreviews
abreview image
Application
Western blot
Sample
Cow Purified protein (purified histone from calf thymus)
Loading amount
1 µg
Specification
purified histone from calf thymus
Gel Running Conditions
Reduced Denaturing (5-20%)
Blocking step
BSA as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: 25°C
Read More

Abcam user community

Verified customer

提交于 Mar 26 2010

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