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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, Dot Blot, WB, ChIP more details
  • Species reactivity
    Reacts with: Mouse, 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 controlWB: 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 ICC/IF: 1/100 - 1/200.
CHIPseq CHIPseq: Use at an assay dependent dilution.
Dot Blot Dot: 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 WB: 1/500. Detects a band of approximately 17 kDa.
ChIP ChIP: Use 2µg for 106 cells.

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 antibodyH3 3 Like Sequence MH921 antibodyH3 3A antibody
      H3 3A antibodyH3 antibodyH3 Histone antibodyH3 Histone Family Member E Pseudogene antibodyH3.4 antibodyH3/A antibodyH3/g antibodyH31_HUMAN antibodyH3F3 antibodyH3F3 antibodyH3FA antibodyH3FT antibodyH3t antibodyHIST1H3J antibodyHIST3H3 antibodyHIST3H3 antibodyHistone cluster 1, H3a antibodyHistone H3 3 Pseudogene antibodyHistone H3.1 antibodyHistone H3.3 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 (acetyl K9) antibody - ChIP Grade images

  • 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.
  • 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.

  • 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

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) »
  • Rajabi H  et al. MUC1-C oncoprotein induces TCF7L2 transcription factor activation and promotes cyclin D1 expression in human breast cancer cells. J Biol Chem 287:10703-13 (2012). ChIP ; Human . Read more (PubMed: 22318732) »

See all 46 Publications for this product

Product Wall

Displaying 1 - 10 of 36 results for Abreviews and Q&A

ChIP

Excellent
Abreviews
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
Username

Verified customer

Submitted Apr 29 2013

It may be worth trying ab10812 since each unit is 100ug, though we have not comopared it to ab4441 in any application. We do not have any more of ab4441 lot GR105501. We have several new lots but we do not test each lot in ChIP. The most recent data we ha...

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Thank you for contacting us.

I can confirm that these antibodies would immunoprecipitate the chromatin irrespective of cell line used, off course if the histone modification is present.

We unfortunately do not offer primer samples howeve...

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Thank you for contacting us. We do not know how these two antibodies compare for ChIP, as they have not been tested against the same samples, to our knowledge. We have not received any complaints for the lot you have, GR105501-1. Antibody ab4441 is the mo...

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Thank you for contacting us. While we do sell positive controls, we do not have these particular controls available in our catalog. If you are looking to test this in Western Blot and have access to Hela cells, we do sell the biochemical sodium butyrate (...

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ChIP

Excellent
Abreviews
Application ChIP
Sample Human Cell lysate - nuclear (t(821) AML cell line)
Specification t(821) AML 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: Formaldehyde
Detection step Real-time PCR
Positive control PU.1 promoter
Negative control IVL
Username

Verified customer

Submitted Nov 12 2012

ChIP

Excellent
Abreviews
Application ChIP
Sample Mouse Cell lysate - other (Mouse ES cells)
Specification Mouse ES cells
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 12 minute(s) and 0 second(s)
Specification of the cross-linking agent: 1% formaldehyde in culture med
Detection step Real-time PCR
Positive control Oct4 promoter
Negative control Chr2 (GAPDH Pseudogene)
Username

Verified customer

Submitted Nov 05 2012

Thank you very much for your call today and for letting us know about the trouble with ab4441 in Western blot.
As we discussed, I'm sending a free of charge vial of ab10812 on the order *** as well as a free of charge vial of the HeLa nuclear lysate ...

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Western blot

Excellent
Abreviews
Application Western blot
Sample Human Cell lysate - nuclear (Hela)
Loading amount 50 µg
Specification Hela
Gel Running Conditions Reduced Denaturing (15% SDS-PAGE)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: RT°C
Username

Verified customer

Submitted Jun 20 2012

Thank you for contacting us.

Antibody ab113674 is the newest antibody of the three and is therefore less thoroughly characterized. Antibodies ab4441 and ab10812 have both been published many times and have received positive customer reviews for...

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