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Anti-Histone H3 (methylated) antibody - ChIP Grade (ab2621)

Abreviews (8)Q&A (17)Specific References (35)

Overview

  • Product nameAnti-Histone H3 (methylated) antibody - ChIP GradeSee all Histone H3 (methylated) primary antibodies ...
  • Description
    Rabbit polyclonal to Histone H3 (methylated) - ChIP Grade
  • SpecificityReacts with Tri-methyl K79 of histone H3. Cross-reactivity to di-methyl K79.
  • Tested applicationsICC/IF, ChIP, WB, ChIP/Chip more details
  • Species reactivity
    Reacts with: Mouse, Rat, Cow, Human, Saccharomyces cerevisiae
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 50 to the C-terminus of Human Histone H3, tri methylated at K79.

    Read Abcam's proprietary immunogen policy

    (Peptide available as ab4557.)

  • Positive controlCalf Thymus Histone Preparation; Hela whole cell extract

Properties

  • FormLiquid
  • Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • Storage bufferpH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: PBS
    Note: 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...
  • PurityImmunogen affinity purified
  • Clonality Polyclonal
  • IsotypeIgG
  • Research Areas

Applications

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.

Application Notes
ICC/IF ICC/IF: Use at an assay dependent concentration.
ChIP ChIP: Use 2-4 µg for 6 µg of chromatin.
WB WB: Use a concentration of 1 µg/ml. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa).Can be blocked with Histone H3 peptide - tri methyl K79 (ab4557).
ChIP/Chip ChIP/Chip: Use at an assay dependent dilution.

Target

  • FunctionCore 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.
  • Sequence similaritiesBelongs to the histone H3 family.
  • Developmental stageExpressed during S phase, then expression strongly decreases as cell division slows down during the process of differentiation.
  • Post-translational
    modifications    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.
  • Cellular localizationNucleus. Chromosome.

    • Target information above from: UniProt accession P68431 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links
    see all
  • Alternative names
      FLJ92264 antibodyH3 histone antibodyH3 histone antibody
      H3 histone family, member A antibodyH3/a antibodyH3/b antibodyH3/c antibodyH3/d antibodyh3/f antibodyH3/h antibodyH3/i antibodyH3/j antibodyH3/k antibodyH3/l antibodyH31_HUMAN antibodyH3F1K antibodyH3F3 antibodyH3F3 antibodyH3FA antibodyH3FB antibodyH3FC antibodyH3FD antibodyH3FF antibodyH3FH antibodyH3FI antibodyH3FJ antibodyH3FK antibodyH3FL antibodyHIST1H3A antibodyHIST1H3B antibodyHIST1H3C antibodyHIST1H3D antibodyHIST1H3E antibodyHIST1H3F antibodyHIST1H3G antibodyHIST1H3H antibodyHIST1H3I antibodyHIST1H3J antibodyHIST3H3 antibodyHIST3H3 antibodyHistone 1, H3a antibodyHistone cluster 1, H3a antibodyHistone cluster 1, H3b antibodyHistone cluster 1, H3c antibodyHistone cluster 1, H3d antibodyHistone cluster 1, H3e antibodyHistone cluster 1, H3f antibodyHistone cluster 1, H3g antibodyHistone cluster 1, H3i antibodyHistone cluster 1, H3j antibodyHistone H 3 antibodyHistone H3.1 antibodyHistone H3.1 antibodyHistone H3/a antibodyHistone H3/b antibodyHistone H3/c antibodyHistone H3/d antibodyHistone H3/f antibodyHistone H3/h antibodyHistone H3/i antibodyHistone H3/j antibodyHistone H3/k antibodyHistone H3/l antibody
    see all

Anti-Histone H3 (methylated) antibody - ChIP Grade images

  • ChIP - Histone H3 (tri methyl K79) antibody - ChIP Grade (ab2621)
    ChIP - Histone H3 (tri methyl K79) antibody - ChIP Grade (ab2621)

    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.

  • Western blot - Histone H3 (tri methyl K79) antibody - ChIP Grade (ab2621)
    Western blot - Histone H3 (tri methyl K79) antibody - ChIP Grade (ab2621)
    All lanes : Anti-Histone H3 (methylated) 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 Histone H3 peptide - unmodified (ab4558) at 0.5 µg/ml
    Lane 3 : Calf Thymus Histone Preparation Nuclear Lysate with Histone H3 peptide - mono methyl K79 (ab4555) at 0.5 µg/ml
    Lane 4 : Calf Thymus Histone Preparation Nuclear Lysate with Histone H3 peptide - di methyl K79 (ab4556) at 0.5 µg/ml
    Lane 5 : Calf Thymus Histone Preparation Nuclear Lysate with Histone H3 peptide - tri methyl K79 (ab4557) at 0.5 µg/ml
    Lane 6 : Calf Thymus Histone Preparation Nuclear Lysate with Histone H4 peptide - di methyl K79 (ab4560) at 0.5 µg/ml
    Lane 7 : Calf Thymus Histone Preparation Nuclear Lysate with Histone H3 peptide - di methyl K9 (ab1772) at 0.5 µg/ml

    Lysates/proteins at 0.5 µg per lane.

    Secondary
    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?)
  • Immunocytochemistry/ Immunofluorescence - Histone H3 (tri methyl K79) antibody - ChIP Grade (ab2621)
    Immunocytochemistry/ Immunofluorescence - Histone H3 (tri methyl K79) antibody - ChIP Grade (ab2621)This image is courtesy of an Abreview submitted by Dr Kirk McManus
    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

  • ChIP - Histone H3 (tri methyl K79) antibody - ChIP Grade (ab2621)
    ChIP - Histone H3 (tri methyl K79) antibody - ChIP Grade (ab2621)This image is a courtesy of Anonymous 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

References for Anti-Histone H3 (methylated) antibody - ChIP Grade (ab2621)

This product has been referenced in:
  • Youn MY  et al. JMJD5, a Jumonji C (JmjC) Domain-containing Protein, Negatively Regulates Osteoclastogenesis by Facilitating NFATc1 Protein Degradation. J Biol Chem 287:12994-3004 (2012). WB ; Human . Read more (PubMed: 22375008) »
  • Lee JS  et al. Codependency of H2B monoubiquitination and nucleosome reassembly on Chd1. Genes Dev 26:914-9 (2012). WB . Read more (PubMed: 22549955) »

See all 35 Publications for this product

Publishing research using ab2621 ? Please let us know so that we can cite the reference in this datasheet

Product Wall

Submit an Abreview for ab2621 Submit a Question for ab2621

Displaying 1 - 10 of 25 results for Abreviews and Q&A

10
Votes: Highest First

I am interested in purchasing this antibody, but I would like to see the QC data for the current lot.

Thank you for your inquiry.

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 AL...

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Hmm, what if the new vial still doesn't work?

Any replacement vial you receive will also be covered by our Abpromise guarantee http://www.abcam.com/abpromise

Hey I just found the PO number. Do I need to provide the address?

Thanks for the PO number. I have located your order in our system.

Just to confirm, would you like to receive another vial of ab2621, a different primary antibody, a credit or refund?

I ordered the antibody through our lab manager and I'm not sure if she can find the PO number. What if we can't find the PO number?

I'm sure we'll be able to find your order somehow.

Please let me know if your lab manager can locate the PO number. If not, it would be helpful to know approximately when you purchased this vial of ab2621.

Thanks!

Thanks for the reply. I have one question: what if the new antibody still doesn't work? Moreover, do you have other H3K79me3 antibody that will be suitable for mouse kidney?

Thanks for your reply.

Regarding your first question, the replacement product is covered by our guarantee just the same as the initial antibody. So if the replacement antibody does not work as expected, we are happy to offer another replacement,...

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Is it the new antibody that is not working or the one that is 2 years old?

Both not working. The old one used to work but now didn't. The new one hasn't worked since it came.

Is it mouse kidney sections that have been tested successf...

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Thanks for your reply.

Although the old vial was purchased too long ago to replace, I am happy to send a vial to replace the most recently purchased vial of ab2621.

1. Please confirm if you would like a new vial of ab2621 as the replac...

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Thank you for the reply and Here are the answers:



1) How long ago did you purchase the while that sudeenly stopped working?

The one suddenly can't work is bought two years ago.






2) ...

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Thanks so much for your reply.

Is it the new antibody that is not working or the one that is 2 years old?

Is it mouse kidney sections that have been tested successfully in the past?

Thanks again for the additional details. I...

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Product code: 2621
Lot number: GR43147-1
Inquiry: It suddenly didn't work in kidney parrafin sections in immunofluorescence. It used to work fine but suddenly lost the efficiency. I've bought a new one with the same lot number, however it didn't...

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Thank you for contacting us.
I'm sorry to hear that the antibody is not working anymore as it used to.

Could you please let me know the following to understand the situation better:

1) How long ago did you purchase the while that...

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ChIP abreview for Anti-Histone H3 (methylated) antibody - ChIP Grade

 Good
Abreviews
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
Read More
abreview image
Username

Verified customer

Submitted Jan 30 2012

Dear Ladies and Gentlemen,
In the future (mid of the year) I want to perform ChIP on Chip experiments. For this purpose I´m planing to use
antibodies from your company which will be verified first in simple ChIP experiments. Our experience...

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Thank you for your enquiry.

Unfortunately 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 g...

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