Anti-Histone H3 (acetyl K9) antibody - ChIP Grade (ab4441)

Overview

  • Product nameAnti-Histone H3 (acetyl K9) antibody - ChIP GradeSee all Histone H3 primary antibodies ...
  • Description
    Rabbit polyclonal to Histone H3 (acetyl K9) - ChIP Grade
  • SpecificityIn Dot blot detects 50ng of mono-acetylated peptide corresponding to position Lys9 in the N-terminal sequence of Histone H3. Does not detect the mono-acetylated peptide corresponding to acetyl-lysine at position 14 or unacetylated Histone H3.
  • Tested applicationsICC/IF, CHIPseq, Flow Cyt, Dot Blot, WB, ChIP, IHC-P more details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Tetrahymena sp., Xenopus laevis, Caenorhabditis elegans, Fruit fly (Drosophila melanogaster), Schizosaccharomyces pombe, Toxoplasma gondii
    Predicted to work with: Saccharomyces cerevisiae
  • Immunogen

    Synthetic peptide: ARTKQTARAcKSTG-C conjugated to KLH, corresponding to amino acids 1-12 of Human Histone H3.

  • Positive control
    • WB: TSA-treated and acid extracted HeLa and NIH/3T3 cells. ICC:polytene chromosomes of Drosophila melanogaster

Properties

Applications

Our Abpromise guarantee covers the use of ab4441 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 1/100 - 1/200.
CHIPseq Use at an assay dependent dilution.
Flow Cyt Use at an assay dependent concentration.
Dot Blot Use a concentration of 0.5 µg/ml. Detects 50ng of mono-acetylated peptide corresponding to position Lys9 in the N-terminal sequence of Histone H3. Does not detect the mono-acetylated peptide corresponding to acetyl-lysine at position 14 or unacetylated Histone H3.
WB 1/500. Detects a band of approximately 17 kDa.
ChIP Use 2µg for 106 cells.
IHC-P Use at an assay dependent concentration.

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
  • 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 - ChIP Grade images

  • Fruit fly (Drosophila melanogaster) Cell (polytene chromosomes) were fixed in formaldehyde, blocked in 1% BSA for 30 minutes and incubated with ab4441 (1/100) for 12 hours.

     

  • 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 ab4441 (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.

  • All lanes : Anti-Histone H3 (acetyl K9) antibody - ChIP Grade (ab4441) at 0.5 µg/ml

    Lane 1 : Untreated HeLa cell acid-extract
    Lane 2 : HeLa cell acid-extract treated with sodium butyrate

    ab4441, Histone H3, acetylated (Lys9) Pab Rabbit x Human

References for Anti-Histone H3 (acetyl K9) antibody - ChIP Grade (ab4441)

This product has been referenced in:
  • Tropberger P  et al. Regulation of transcription through acetylation of H3K122 on the lateral surface of the histone octamer. Cell 152:859-72 (2013). ChIP ; Human . Read more (PubMed: 23415232) »
  • Wolf G  et al. Epigenetic marking and repression of porcine endogenous retroviruses. J Gen Virol 94:960-70 (2013). ChIP . Read more (PubMed: 23324470) »

See all 71 Publications for this product

Product Wall

Application Western blot
Loading amount 1 µg
Gel Running Conditions Reduced Denaturing (18 % SDS-PAGE)
Sample Dictyostelium discoideum Purified protein (Vegetative cells)
Specification Vegetative cells
Blocking step No blocking step used for 30 minute(s) · Concentration: 100% · Temperature: 25°C
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Submitted Mar 31 2014

Application Flow Cytometry
Fixation Paraformaldehyde
Permeabilization Yes - eBioscience Permeabilization Buffer
Sample Human Cell (Differentiated Haematopoietic Stem Cells)
Specification Differentiated Haematopoietic Stem Cells
Gating Strategy Isotype negative control (white)
Preparation Cell harvesting/tissue preparation method: Typsin-EDTA Cell Dissociation
Sample buffer: PBS and 10% FBS
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Submitted Feb 27 2014

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing
Sample Human Tissue lysate - whole (Patient Dermal Fibroblasts)
Specification Patient Dermal Fibroblasts
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
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Submitted Feb 01 2014

Application Western blot
Loading amount 10 µg
Gel Running Conditions Reduced Denaturing (12)
Sample Human Cell lysate - nuclear (Human HeLa cell)
Specification Human HeLa cell
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
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Submitted Nov 05 2013

Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing (12)
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
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Submitted Oct 24 2013

Application Western blot
Loading amount 3 µg
Gel Running Conditions Reduced Denaturing (12)
Sample Fruit fly (Drosophila melanogaster) Tissue lysate - nuclear (Fruit fly embryo)
Specification Fruit fly embryo
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
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Submitted Oct 21 2013

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step Serum as blocking agent for 45 minute(s) · Concentration: 1.5% · Temperature: 25°C
Antigen retrieval step Enzymatic
Sample Rat Tissue sections (GONAD)
Specification GONAD
Permeabilization No
Fixative Formaldehyde
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Submitted Sep 20 2013

Application ChIP
Detection step Real-time PCR
Sample Human Cell lysate - other (epithelial cells)
Specification epithelial cells
Negative control rIgG
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
Positive control input DNA
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Submitted Sep 12 2013


For ab4441 we test on the following primers:

Active
GADPH: ab83588 (f), ab83589 (r)
RPL30: ab74244 (f), ab83296 (r)
ALDOA: ab83591 (f), ab83592 (r)

Inactive
Myo-D: ab83600 (f), ab83601 (r)
Serpina: ab83603 (f)...

Read More
Application ChIP
Sample Mouse Cell lysate - whole cell (C2C12 mouse myoblast cell line)
Specification C2C12 mouse myoblast cell line
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: 0.75% formaldehyde
Detection step Real-time PCR
Positive control Unexposed C2C12 cells at day 2 of differentiation
Negative control Beads only and mouse IgG
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Submitted Apr 29 2013

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