Anti-Histone H3 (acetyl K9) antibody [AH3-120] - ChIP Grade (ab12179)

Overview

  • Product nameAnti-Histone H3 (acetyl K9) antibody [AH3-120] - ChIP GradeSee all Histone H3 primary antibodies ...
  • Description
    Mouse monoclonal [AH3-120] to Histone H3 (acetyl K9) - ChIP Grade
  • Tested applicationsFlow Cyt, IHC-P, ChIP, WB, ELISA, ICC, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Human, Arabidopsis thaliana, Fruit fly (Drosophila melanogaster), Rice
    Predicted to work with: Rat, Chicken, Cow, Xenopus laevis, Caenorhabditis elegans, a wide range of other species
  • Immunogen

    Synthetic peptide corresponding to Histone H3 aa 7-20 (N terminal). aa 7 to 20 and Ac-Lys9.

  • Positive control
    • IHC-P: Human normal colon FFPE tissue sections.
  • General notesStorage in frost-free freezers is not recommended. If slight turbidity occurs upon prolonged storage, clarify the solution by centrifugation before use. Working dilutions should be discarded if not used within 12 hours.

Properties

Applications

Our Abpromise guarantee covers the use of ab12179 in the following tested applications.

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

Application Abreviews Notes
Flow Cyt Use at an assay dependent concentration.

(PMID 19584087)

IHC-P Use a concentration of 10 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ChIP Use 5-10 µg for 25 µg of chromatin.
WB Use a concentration of 1 - 2 µg/ml. Detects a band of approximately 17 kDa.
ELISA Use at an assay dependent concentration.
ICC Use at an assay dependent concentration.
ICC/IF 1/1000.

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.
  • Information by UniProt
  • Database links
  • Alternative names
    • H 3 antibody
    • H3 3 Like Sequence MH921 antibody
    • H3 3A antibody
    • H3 3A antibody
    • H3 antibody
    • H3 Histone antibody
    • H3 Histone Family Member E Pseudogene antibody
    • H3.4 antibody
    • H3/A antibody
    • H3/g antibody
    • H31_HUMAN antibody
    • H3F3 antibody
    • H3F3 antibody
    • H3FA antibody
    • H3FT antibody
    • H3t antibody
    • HIST1H3J antibody
    • HIST3H3 antibody
    • HIST3H3 antibody
    • Histone cluster 1, H3a antibody
    • Histone H3 3 Pseudogene antibody
    • Histone H3.1 antibody
    • Histone H3.3 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

Anti-Histone H3 (acetyl K9) antibody [AH3-120] - ChIP Grade images

  • IHC image of ab12179 staining Histone H3 (acetyl K9) in human colon formalin fixed paraffin embedded tissue sections, performed on a Leica Bond. 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 ab12179, 10µg/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. No primary antibody was used in the negative control (shown on the inset).

    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.

  • SK-N-SH cells fixed in 4% paraformaldehyde, permeabilized in 0.5% Triton X-100 and incubated for 1 hour with ab12179 (1/1000 dilution). ab12179 staining is localized to the nucleus (red). The cells were counterstained with DAPI (blue). 100x magnification.

  • 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  ab12179 (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.
        

  • ab12179 at a 1/1000 dilution staining Stylonychia lemnae (single cell organism, transcriptionally active macronucleus) by ICC/IF. The cells were paraformaldehyde fixed and incubated with the antibody for 12 hours. An Alexa Fluor ® 488 conjugated goat anti-mouse IgG antibody was used as the secondary.

    In the image Histone H3 (acetyl K9) staining is red and is found in macronuclei only. Micronuclei remain unstained and are shown in blue (nucleic acid counterstain). Alpha tubulin is also stained (green).

    See Abreview

References for Anti-Histone H3 (acetyl K9) antibody [AH3-120] - ChIP Grade (ab12179)

This product has been referenced in:
  • Wang M  et al. EGFR-mediated chromatin condensation protects KRAS-mutant cancer cells against ionizing radiation. Cancer Res 74:2825-34 (2014). WB ; Human . Read more (PubMed: 24648348) »
  • Dong X  et al. Modification of histone acetylation facilitates hepatic differentiation of human bone marrow mesenchymal stem cells. PLoS One 8:e63405 (2013). ICC/IF ; Human . Read more (PubMed: 23658825) »

See all 12 Publications for this product

Product Wall

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement.

This shipment is expected to arrive Tuesday, August 21 2012. To check the...

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Thank you for your question.

I hope I can clarify the protocol:

Step 5. is a repetition of step 4.: this provides you the pellet for furtheruse.
Step 6. of the protocol: Re-suspend the pellet in 0.2 N HCl at a density of 4x107 ...

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Thank you for your reply.

I am sorry for the misunderstanding, I assumed that you performed an IP.

As Histones can be a bit tricky, we usually suggest using a nuclear fraction lysate or to follow our Histone extraction protocol (see...

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I am sorry to hear you are experiencing difficulties with our products. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

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Application ChIP
Sample Rice Cell lysate - whole cell (14 days old seedlings grown under dark condition)
Specification 14 days old seedlings grown under dark condition
Type Native ChIP (N-ChIP)
Detection step Real-time PCR
Positive control Gene of interest
Negative control No Antibody control
Username

Mr,Mr,Mr. Yutaka Sato

Verified customer

Submitted Dec 07 2007

Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (MEFs SV40 transformed)
Specification MEFs SV40 transformed
Fixative Paraformaldehyde
Permeabilization Yes - 0.5% triton 10 min RT
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: RT°C
Username

Mr. Mark Rochman

Verified customer

Submitted Aug 13 2007

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Stylonychia lemnae Cell (single cell organism)
Specification single cell organism
Fixative Paraformaldehyde
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 4%
Username

Dr. Jan Postberg

Verified customer

Submitted Dec 12 2006

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Arabidopsis thaliana Cell (Landsberg culture cell)
Specification Landsberg culture cell
Fixative Paraformaldehyde
Blocking step BSA as blocking agent for 1 hour(s) and 40 minute(s) · Concentration: 3 %
Username

Dr. Peter McKeown

Verified customer

Submitted Nov 16 2006

Thank you for your enquiry and I'm sorry to hear that you are experiencing difficulty with ab12179. At this time I would like to make a few suggestions to try to help you out. First, I recommend running a positive control. One recommendation is calf th...

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