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Epigenetics and Nuclear Signaling Histones H2B Methylated
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Anti-Histone H2B (mono methyl K5) antibody - 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

You may also be interested in

Secondary
Product image
Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
Protein
Product image
Recombinant Human Histone H2B protein (ab92409)

View more associated products

Overview

  • Product name

    Anti-Histone H2B (mono methyl K5) antibody - ChIP Grade
    See all Histone H2B primary antibodies
  • Description

    Rabbit polyclonal to Histone H2B (mono methyl K5) - ChIP Grade
  • Host species

    Rabbit
  • Specificity

    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.
  • Tested Applications & Species

    Application Species
    ChIP
    Human
    IHC-P
    Human
    IP
    Human
    WB
    Cow
    See all applications and species data
  • Immunogen

    Synthetic peptide within Human Histone H2B aa 1-100 (mono methyl K5) conjugated to keyhole limpet haemocyanin. The exact sequence is proprietary.
    (Peptide available as ab13211)

Properties

  • Form

    Liquid
  • Storage instructions

    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.
  • Storage buffer

    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...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

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

Associated products

  • 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)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab12929 in the following tested applications.

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

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
ChIP
Human
IHC-P
Human
IP
Human
WB
Cow
All applications
Mouse
Rat
Chicken
Xenopus laevis
Application Abreviews Notes
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.
Notes
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.

Target

  • Relevance

    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.
  • Cellular localization

    Nuclear
  • Database links

    • Entrez Gene: 337874 Human
    • Entrez Gene: 54145 Human
    • Entrez Gene: 8349 Human
    • Entrez Gene: 104021 Mouse
    • Entrez Gene: 64647 Rat
    • 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
  • Alternative names

    • 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

Images

  • 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.

Protocols

  • PCR primers used to test ab12929 in ChIP at Abcam

Click here to view the general protocols

Datasheets and documents

    • Datasheet
  • References (4)

    Publishing research using ab12929? Please let us know so that we can cite the reference in this datasheet.

    ab12929 has been referenced in 4 publications.

    • 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

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    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

    Submitted 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

    Submitted 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

    Submitted 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

    Submitted Jan 08 2014

    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

    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

    Submitted Feb 15 2010

    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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