Anti-Histone H2B抗体[mAbcam 52484] - ChIP Grade (ab52484)

概述

  • 产品名称
    Anti-Histone H2B抗体[mAbcam 52484] - ChIP Grade
    参阅全部 Histone H2B 一抗
  • 描述
    小鼠单克隆抗体[mAbcam 52484] to Histone H2B - ChIP Grade
  • 经测试应用
    适用于: IP, Flow Cyt, WB, ICC/IF, ChIP, IHC-P, IHC-Frmore details
  • 种属反应性
    与反应: Mouse, Rat, Chicken, Human, Drosophila melanogaster, Zebrafish
    预测可用于: Cow, Xenopus laevis, Caenorhabditis elegans, Orangutan
  • 免疫原

    Synthetic peptide corresponding to Human Histone H2B aa 100 to the C-terminus (C terminal) conjugated to Keyhole Limpet Haemocyanin (KLH).

  • 阳性对照
    • This antibody gave a positive signal in NIH3T3 and PC12 whole cell lysates and when tested against Histone H2B recombinant protein.
  • 常规说明

    This antibody clone is manufactured by Abcam.

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 or -80°C. Avoid freeze / thaw cycle.
  • 存储溶液
    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: PBS

    Please note that some batches of ab52484 may contain 0.4M arginine. Please contact Scientific Support for further information.
  • Concentration information loading...
  • 纯度
    IgG fraction
  • 克隆
    单克隆
  • 克隆编号
    mAbcam 52484
  • 骨髓瘤
    Sp2/0
  • 同种型
    IgG1
  • 研究领域

应用

Our Abpromise guarantee covers the use of ab52484 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
IP Use a concentration of 5 µg/ml.
Flow Cyt Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, 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).
ICC/IF Use a concentration of 1 µg/ml.
ChIP Use 5 µg for µg of chromatin.
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IHC-Fr Use at an assay dependent concentration.

靶标

  • 相关性
    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.
  • 细胞定位
    Nuclear
  • 数据库链接
  • 别名
    • 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
    • HIRIP2 antibody
    • HIST1H2BB antibody
    • HIST1H2BD antibody
    • HIST1H2BH antibody
    • HIST1H2BL antibody
    • HIST1H2BM antibody
    • HIST1H2BN antibody
    • HIST2H2BE antibody
    • Histone H2B 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 protein antibody
    see all

Anti-Histone H2B antibody [mAbcam 52484] - ChIP Grade 图像

  • Chromatin was prepared from HeLa cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10min. The  ChIP was performed with 25µg of chromatin, 5µg of  ab52484 (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). Primers and probes are located in the first kb of the transcribed region.  

  • ab52484 staining Histone H2B in Fruit fly (Drosophila melanogaster) embryo cells by ICC/IF (Immunocytochemistry/immunofluorescence). Embryos were washed in 1x PBS + 0.1% Trition, cells were fixed with heat, and blocked with 10% BSA for 12 hours at 4°C. Samples were incubated with primary antibody (1/1000) for 24 hours. An Alexa Fluor®594-conjugated Rabbit anti-mouse IgG polyclonal (1/5000) was used as the secondary antibody.

    See Abreview

  • All lanes : Anti-Histone H2B antibody [mAbcam 52484] - ChIP Grade (ab52484) at 5 µg/ml

    Lane 1 : Histone H1 recombinant protein.
    Lane 2 : Histone H2A recombinant protein.
    Lane 3 : Histone H2B recombinant protein.
    Lane 4 : Histone H3 recombinant protein.
    Lane 5 : Histone H4 recombinant protein.

    Lysates/proteins at 0.1 µg per lane.

    Secondary
    Rabbit polyclonal to Mouse IgG - H&L (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?)
  • ICC/IF image of ab52484 stained human HeLa cells. The cells were 4% PFA fixed (10 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab52484, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).

  • IHC image of Histone H2B staining in human breast carcinoma FFPE section, performed on a BondTM 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 ab52484, 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.
  • All lanes : Anti-Histone H2B antibody [mAbcam 52484] - ChIP Grade (ab52484) at 5 µg/ml

    Lane 1 : Marker
    Lane 2 : Zebrafish brain homogenate at 20 µg
    Lane 3 : Zebrafish heart homogenate at 10 µg
    Lane 4 : Zebrafish liver homogenate at 10 µg
    Lane 5 : Zebrafish skeletal muscle homogenate at 10 µg
    Lane 6 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

    Secondary
    Goat polyclonal to Mouse IgG – H&L – Pre-Adsorbed (HRP) at 1/6000 dilution
    Developed using the ECL technique

    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 : 1 minute
  • Overlay histogram showing HeLa cells stained with ab52484 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab52484, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

  • All lanes : Anti-Histone H2B antibody [mAbcam 52484] - ChIP Grade (ab52484) at 1 µg/ml

    Lane 1 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lane 2 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate

    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 : 17 kDa (why is the actual band size different from the predicted?)


    Exposure time : 1 minute
  • Histone H2B was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Mouse monoclonal to Histone H2B 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 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab52484.
    Secondary: Protein G-HRP at 1/500 dilution.
    Band: 17kDa; Histone H2B; non specific bands - 26kDa: We are unsure as to the identity of this extra band.

Anti-Histone H2B antibody [mAbcam 52484] - ChIP Grade (ab52484)参考文献

This product has been referenced in:
  • Bui M  et al. Internal modifications in the CENP-A nucleosome modulate centromeric dynamics. Epigenetics Chromatin 10:17 (2017). WB ; Human . Read more (PubMed: 28396698) »
  • Sawhney S  et al. Alpha-enolase is upregulated on the cell surface and responds to plasminogen activation in mice expressing a ?133p53a mimic. PLoS One 10:e0116270 (2015). WB ; Mouse . Read more (PubMed: 25643152) »

See all 7 Publications for this product

Product Wall

Application
ChIP
Sample
Mouse Cell lysate - whole cell (Mouse colon cells)
Specification
Mouse colon cells
Detection step
Real-time PCR
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: Formaldehyde 11%
Username

Abcam user community

Verified customer

提交于 Jan 30 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (Colon)
Specification
Colon
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 20%
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

提交于 Sep 06 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Breast)
Specification
Breast
Blocking step
(agent) for 1 hour(s) and 0 minute(s) · Concentration: 20%
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

提交于 Aug 26 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 4°C
Sample
Fruit fly (Drosophila melanogaster) Cell (Drosophila Embryo)
Specification
Drosophila Embryo
Permeabilization
No
Fixative
Heat
Username

Abcam user community

Verified customer

提交于 Jul 09 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - whole cell (Mouse lung)
Loading amount
100000 cells
Specification
Mouse lung
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
Username

Abcam user community

Verified customer

提交于 Nov 22 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
ChIP
Sample
Human Cell lysate - whole cell (Breast cell line)
Specification
Breast cell line
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 0 second(s)
Detection step
Real-time PCR
Username

Abcam user community

Verified customer

提交于 Nov 20 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (Breast cell line)
Loading amount
100000 cells
Specification
Breast cell line
Gel Running Conditions
Non-reduced Denaturing (12.5%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
Username

Abcam user community

Verified customer

提交于 Nov 20 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Chicken Cell lysate - nuclear (QT6 cells)
Loading amount
7.5 µg
Specification
QT6 cells
Gel Running Conditions
Reduced Denaturing (12% acrylamide)
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Nov 14 2012

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Specification
HeLa
Fixative
Methanol
Username

Dr. Kirk Mcmanus

Verified customer

提交于 May 09 2008

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

注册