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  1. Link

    histone-h3-antibody-nuclear-marker-and-chip-grade-ab1791.pdf

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Isotype/Loading Controls Loading Controls Histone H3
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Anti-Histone H3 antibody - Nuclear Marker and ChIP Grade (ab1791)

  • Datasheet
  • SDS
Reviews (232)Q&A (49)References (3819)

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Western blot - Anti-Histone H3 antibody - Nuclear Marker and ChIP Grade (ab1791)
  • Immunoprecipitation - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • Immunocytochemistry - Anti-Histone H3 antibody - Nuclear Marker and ChIP Grade (ab1791)
  • Western blot - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • Immunocytochemistry - Anti-Histone H3 antibody - Nuclear Marker and ChIP Grade (ab1791)
  • Western blot - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • Western blot - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)

Key features and details

  • Rabbit polyclonal to Histone H3 - Nuclear Marker and ChIP Grade
  • Suitable for: IHC-P, ChIP, IP, WB, ICC/IF
  • Reacts with: Mouse, Rat, Human, Saccharomyces cerevisiae, Xenopus laevis, Arabidopsis thaliana, Drosophila melanogaster, Indian muntjac, Schizosaccharomyces pombe
  • Isotype: IgG

Get better batch-to-batch reproducibility with a recombinant antibody

Product image
Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (ab176842)
  • Research with confidence – consistent and reproducible results with every batch
  • Long-term and scalable supply – powered by recombinant technology for fast production
  • Success from the first experiment – confirmed specificity through extensive validation
  • Ethical standards compliant – production is animal-free

Overview

  • Product name

    Anti-Histone H3 antibody - Nuclear Marker and ChIP Grade
    See all Histone H3 primary antibodies
  • Description

    Rabbit polyclonal to Histone H3 - Nuclear Marker and ChIP Grade
  • Host species

    Rabbit
  • Specificity

    Based only on sequence homology, we expect the antibody to react with multiple variants of H3 such as H3.1, H3.2 and H3.3.

  • Tested applications

    Suitable for: IHC-P, ChIP, IP, WB, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human, Saccharomyces cerevisiae, Xenopus laevis, Arabidopsis thaliana, Drosophila melanogaster, Indian muntjac, Schizosaccharomyces pombe
    Predicted to work with: Chicken, Dog, Caenorhabditis elegans, Ferret, Zebrafish, a wide range of other species, Mammals, Silk worm, Dictyostelium discoideum, Rainbow trout, Neurospora crassa, Toxoplasma gondii, Rice, Schistosoma mansoni, Candida albicans, Cyanidioschyzon merolae
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
    (Peptide available as ab12149)

  • General notes

    A recombinant rabbit monoclonal alternative is available to this target – ab176842

    Rabbit polyclonal IgG (ab171870) is suitable for use as an isotype control with this antibody.

     

     

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

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

    Preservative: 0.02% Sodium azide
    Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

    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

    • Isotype/Loading Controls
    • Loading Controls
    • Histone H3

Associated products

  • Alternative Versions

    • HRP Anti-Histone H3 antibody - Nuclear Loading Control (ab21054)
  • ChIP Related Products

    • Rabbit Anti-Mouse IgG H&L (ab46540)
    • ChIP Kit (ab500)
  • Coimmunogen

    • Anti-Histone H3 antibody - Nuclear Marker and ChIP Grade (ab1791)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Immunizing Peptide (Blocking)

    • Human Histone H3 peptide (ab12149)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Recombinant Protein

    • Recombinant human Histone H3 protein (Active) (ab198757)
  • Related Products

    • Histone H3 Total Quantification Kit (Colorimetric) (ab115091)
    • Prestained Protein Ladder – Broad molecular weight (10-245 kDa) (ab116028)
    • Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220)
    • Human Histone H3 (tri methyl K4) peptide (ab1342)
    • Human Histone H3 (tri methyl K9) peptide (ab1773)
    • Human Histone H3 (tri methyl K27) peptide (ab1782)
    • Histone H3 Modification Multiplex Assay Kit (Colorimetric) (ab185910)
    • Anti-Histone H3 antibody [mAbcam 24834] - Nuclear Loading Control and ChIP Grade (ab24834)
    • Anti-Histone H3 (tri methyl K27) antibody [mAbcam 6002] - ChIP Grade (ab6002)
    • Human Histone H3 (unmodified) peptide (ab7228)
    • Human Histone H3 (di methyl K4) peptide (ab7768)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab1791 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
IHC-P (20)
1/100 - 1/400. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ChIP (46)
Use 2µg for 106 cells.
IP (4)
Use a concentration of 5 µg/ml.
WB (121)
1/1000 - 1/5000. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa).Can be blocked with Human Histone H3 peptide (ab12149).

We recommend Goat Anti-Rabbit IgG H&L (HRP) (ab6721) secondary antibody.

ICC/IF (27)
Use a concentration of 1 µg/ml.
Notes
IHC-P
1/100 - 1/400. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ChIP
Use 2µg for 106 cells.
IP
Use a concentration of 5 µg/ml.
WB
1/1000 - 1/5000. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa).Can be blocked with Human Histone H3 peptide (ab12149).

We recommend Goat Anti-Rabbit IgG H&L (HRP) (ab6721) secondary antibody.

ICC/IF
Use a concentration of 1 µg/ml.

Target

  • Function

    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.
  • Sequence similarities

    Belongs to the histone H3 family.
  • Developmental stage

    Expressed 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 localization

    Nucleus. 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

    • Entrez Gene: 8350 Human
    • Entrez Gene: 8351 Human
    • Entrez Gene: 8352 Human
    • Entrez Gene: 8353 Human
    • Entrez Gene: 8354 Human
    • Entrez Gene: 8355 Human
    • Entrez Gene: 8356 Human
    • Entrez Gene: 8357 Human
    • Entrez Gene: 8358 Human
    • Entrez Gene: 8968 Human
    • Entrez Gene: 319152 Mouse
    • Entrez Gene: 319153 Mouse
    • Entrez Gene: 360198 Mouse
    • Entrez Gene: 97908 Mouse
    • Entrez Gene: 100364501 Rat
    • Entrez Gene: 100365669 Rat
    • Entrez Gene: 291159 Rat
    • Entrez Gene: 679994 Rat
    • Entrez Gene: 680511 Rat
    • Entrez Gene: 682330 Rat
    • Omim: 602810 Human
    • SwissProt: P84229 Chicken
    • SwissProt: P68431 Human
    • SwissProt: P68433 Mouse
    • SwissProt: Q6LED0 Rat
    • Unigene: 132854 Human
    • Unigene: 247813 Human
    • Unigene: 247814 Human
    • Unigene: 248176 Human
    • Unigene: 443021 Human
    • Unigene: 484990 Human
    • Unigene: 532144 Human
    • Unigene: 533292 Human
    • Unigene: 546315 Human
    • Unigene: 586261 Human
    • Unigene: 591778 Human
    • Unigene: 221301 Mouse
    • Unigene: 261657 Mouse
    • Unigene: 377874 Mouse
    • Unigene: 390558 Mouse
    • Unigene: 397328 Mouse
    • Unigene: 138090 Rat
    see all
  • Alternative names

    • H3 histone family member E pseudogene antibody
    • H3 histone family, member A antibody
    • H3/A antibody
    • H31_HUMAN antibody
    • H3F3 antibody
    • H3FA antibody
    • Hist1h3a antibody
    • HIST1H3B antibody
    • HIST1H3C antibody
    • HIST1H3D antibody
    • HIST1H3E antibody
    • HIST1H3F antibody
    • HIST1H3G antibody
    • HIST1H3H antibody
    • HIST1H3I antibody
    • HIST1H3J antibody
    • HIST3H3 antibody
    • histone 1, H3a antibody
    • Histone cluster 1, H3a antibody
    • Histone H3 3 pseudogene antibody
    • Histone H3.1 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

Images

  • Western blot - Anti-Histone H3 antibody - Nuclear Marker and ChIP Grade (ab1791)
    Western blot - Anti-Histone H3 antibody - Nuclear Marker and ChIP Grade (ab1791)
    All lanes : Anti-Histone H3 antibody - Nuclear Marker and ChIP Grade (ab1791) at 1/1000 dilution

    Lane 1 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
    Lane 3 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
    Lane 4 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate with Human Histone H3 peptide (ab12149) at 1 µg/ml
    Lane 5 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate with Human Histone H3 peptide (ab12149) at 1 µg/ml
    Lane 6 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate with Human Histone H3 peptide (ab12149) at 1 µg/ml

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 15 kDa
    Observed band size: 17 kDa why is the actual band size different from the predicted?


    Exposure time: 10 seconds


    This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab1791 overnight at 4°C.

    Goat Anti-Rabbit IgG H&L (HRP) (ab6721) secondary antibody was used for detection.

    Antibody binding was visualised using ECL development solution ab133406.

  • Immunoprecipitation - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
    Immunoprecipitation - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)

    Histone H3 - ChIP Grade was immunoprecipitated using 0.5mg HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell extract, 5 µg of Rabbit polyclonal to and 50 µl of protein G magnetic beads (+). No antibody was added to the control (-).

    The antibody was incubated under agitation with Protein G beads for 10 minutes, HeLa whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10 minutes under agitation.

    Proteins were eluted by addition of 40 µl SDS loading buffer and incubated for 10 minutes at 70°C; 10 µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab1791.

    Secondary Antibody: Mouse anti-rabbit HRP light chain (HRP) (ab99697).

    Band: 15kDa; Histone H3 - ChIP Grade

  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
    ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)

    Chromatin from Xenopus laevis oocytes was prepared according to the Abcam X-ChIP protocol.

    Oocytes were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25 mg of chromatin, 3 mg of ab7834 (anti-H3, light blue) and 3 µg of ab1791 (anti-H3, dark blue), and 20 ml of Protein A/G sepharose beads. A non-specific antibody was used as a control (yellow).

    The immunoprecipitated DNA was quantified by real time PCR (Taqman approach).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)This image is courtesy of an anonymous Abreview

    ab1796 staining Histone H3 in mouse liver tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections).

    Tissue was fixed with paraformaldehyde, permeabilized with 0.05% Triton X-100 in PBS for 30 minutes and blocked with 5% BSA for 1 hour; antigen retrieval was by heat mediation in sodium citrate pH 6. Samples were incubated with the primary antibody (1/500 in blocking buffer) for 16 hours at 4°C. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG polyclonal (1/400) was used as the secondary antibody.

    See Abreview

  • Immunocytochemistry - Anti-Histone H3 antibody - Nuclear Marker and ChIP Grade (ab1791)
    Immunocytochemistry - Anti-Histone H3 antibody - Nuclear Marker and ChIP Grade (ab1791)

    ab1791 staining Histone H3 in HeLa cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab1791 at 0.1 µg/mL and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

  • Western blot - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
    Western blot - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)This image is courtesy of an anonymous Abreview
    All lanes : Anti-Histone H3 antibody - Nuclear Marker and ChIP Grade (ab1791) at 1/1000 dilution

    Lane 1 : Mouse skeletal muscle mitochondrial fraction
    Lane 2 : Mouse skeletal muscle nuclear fraction

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : HRP-conjugated goat anti-rabbit IgG at 1/4000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 15 kDa
    Observed band size: 17 kDa why is the actual band size different from the predicted?


    Exposure time: 7 minutes


    Blocked with 3% milk for 1 hour at 25°C.

    Incubated with the primary antibody for 16 hours at 4°C in 3% milk in TBS-tween.

    See Abreview

  • Immunocytochemistry - Anti-Histone H3 antibody - Nuclear Marker and ChIP Grade (ab1791)
    Immunocytochemistry - Anti-Histone H3 antibody - Nuclear Marker and ChIP Grade (ab1791)This image is courtesy of an anonymous Abreview

    ab1791 staining Histone H3 in HeLa (Human epithelial cell line from cervix adenocarcinoma) by ICC/IF (Immunocytochemistry/immunofluorescence).

    Cells were fixed with methanol and blocked with 0.2% fish scale gelatin for 1 hour at 25°C. Samples were incubated with the primary antibody (1/300 in PBS + 0.2% gelatin) for 20 minutes at 25°C. An Alexa Fluor® 488-conjugated donkey anti-rabbit IgG polyclonal (1/500) was used as the secondary antibody.

    Green - Histone H3.
    Blue - DAPI.
    Red - Tubulin.

    See Abreview

  • Western blot - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
    Western blot - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
    All lanes : Anti-Histone H3 antibody - Nuclear Marker and ChIP Grade (ab1791) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : NIH/3T3 whole cell lysate (ab7179)
    Lane 3 : Drosophila embryo nuclear extract (from melanogaster embryos 0-12Hr)
    Lane 4 : S.cerevisiae (Y190) Whole Cell Lysate
    Lane 5 : S.pombe Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 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?



    ab1791 is tested in western blot on a range of species.  We recommend loading higher amounts of protein (20-30ug) to increase the signal in yeast lysates
  • Western blot - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
    Western blot - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)This image is courtesy of John E. Mueller and J. Ruth German (Mary Bryk lab)

    Rabbit polyclonal to Histone H3 (ab1791) at 1/5000 on S. cerevisiae whole cell lysate (40 ug per lane).

    Protein resolved on 15% SDS-PAGE gel. After transfer to PVDF membrane, blots were blocked in 1X PBS, 0.1% Tween-20, and 5% milk. ab1791 was diluted in 5 ml blocking buffer at 1/5000.  Blots plus primary antibodies were either incubated overnight at 4°C or at RT for 2 hours. Blots were washed 6X for 10 minutes each in PBS with 0.1% Tween-20 before addition of secondary antibodies. Secondary antibodies were diluted 1/2,000 in blocking buffer and incubated with blots for 2 hours at RT. Secondary blots were washed 4X for 10 minutes each in PBS with 0.1% Tween-20 and 2X for 10 minutes each in PBS.

  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
    ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)

    Chromatin was prepared from HeLa (Human epithelial cell line from cervix adenocarcinoma) cells according to the Abcam X-ChIP protocol.

    Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25 µg of chromatin, 2 µg of ab1791 (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 for active and inactive loci, Sybr green approach for heterochromatic loci). Primers and probes are located in the first kb of the transcribed region.

  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
    ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)

    The ChIP was performed with chromatin from mouse gut cell lysate and ab1791 at 1/250 dilution.

    Negative control: No antibody was used (right bar).

    The immunoprecipitated DNA was quantified by real time PCR.

     

    See Abreview

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)

    Paraffin-embedded rat brain tissue stained for Histone H3 using ab1791 at 1/8000 dilution in immunohistochemical analysis.

    See Abreview

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)This image is courtesy of an anonymous Abreview

    ab1791 staining Histone H3 in human infantile fibromatosis tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections).

    Tissue was fixed with formaldehyde and blocked with 1% FBS/BSA for 3 hours at room temperature; antigen retrieval was by heat mediation in Tris pH 9. Samples were incubated with primary antibody (1/100 in TBS + 1% BSA + 1% FBS) for 16 hours. An un-diluted HRP-conjugated goat anti-rabbit IgG polyclonal was used as the secondary antibody.

    See Abreview

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)

    ab1791 staining Histone H3 (red) in rat brain tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections).

    Tissue was fixed with formaldehyde, permeabilized with 0.1% TBS-TritonX and blocked with 10% serum for 1 hour at 25°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with the primary antibody (1/500 in 10% normal goat serum) for 24 hours at 24°C. An Alexa Fluor® 594-conjugated goat anti-rabbit IgG polyclonal (1/500) was used as the secondary antibody.

    Green - Nucleus staining.
    Red - Histone H3 staining.

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (3819)

Publishing research using ab1791? Please let us know so that we can cite the reference in this datasheet.

ab1791 has been referenced in 3819 publications.

  • Yuan L  et al. The transcriptional repressors VAL1 and VAL2 recruit PRC2 for genome-wide Polycomb silencing in Arabidopsis. Nucleic Acids Res 49:98-113 (2021). PubMed: 33270882
  • Aguilar R  et al. Epigenetic silencing of the osteoblast-lineage gene program during hippocampal maturation. J Cell Biochem 122:367-384 (2021). PubMed: 33135214
  • Zhang Z  et al. Global H3K79 di-methylation mediates DNA damage response to PAH exposure in Chinese coke oven workers. Environ Pollut 268:115956 (2021). PubMed: 33158619
  • Muniz L  et al. Circular ANRIL isoforms switch from repressors to activators of p15/CDKN2B expression during RAF1 oncogene-induced senescence. RNA Biol 18:404-420 (2021). PubMed: 32862732
  • Mayer BF & Charron JB Transcriptional memories mediate the plasticity of cold stress responses to enable morphological acclimation in Brachypodium distachyon. New Phytol 229:1615-1634 (2021). PubMed: 32966623
View all Publications for this product

Customer reviews and Q&As

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1-10 of 281 Abreviews or Q&A

Immunocytochemistry/ Immunofluorescence abreview for Anti-Histone H3 antibody - Nuclear Marker and ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HUVEC)
Permeabilization
No
Specification
HUVEC
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0.1% · Temperature: RT°C
Fixative
Methanol
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Mr. Łukasz Witucki

Verified customer

Submitted Jun 02 2021

Immunocytochemistry/ Immunofluorescence abreview for Anti-Histone H3 antibody - Nuclear Marker and ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (CRC cell line)
Permeabilization
No
Specification
CRC cell line
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative
Acetone
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted May 21 2021

Western blot abreview for Anti-Histone H3 antibody - Nuclear Marker and ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Rat Cell lysate - nuclear (Brain)
Gel Running Conditions
Reduced Denaturing (4-15% precast gel)
Loading amount
10 µg
Treatment
beta-mercaptoethanol
Specification
Brain
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

MISS. Francesca Mcewan

Verified customer

Submitted Apr 28 2021

Western blot abreview for Anti-Histone H3 antibody - Nuclear Marker and ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Mouse Tissue lysate - whole (Heart)
Gel Running Conditions
Reduced Denaturing (4-12%)
Loading amount
30 µg
Specification
Heart
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Aug 13 2020

Western blot abreview for Anti-Histone H3 antibody - Nuclear Marker and ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (cell line)
Gel Running Conditions
Reduced Denaturing (12)
Loading amount
100 µg
Specification
cell line
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Oct 28 2021

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Histone H3 antibody - Nuclear Marker and ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Specification
Brain
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative
Formaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

MR. Carl Hobbs

Verified customer

Submitted Oct 24 2021

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Histone H3 antibody - Nuclear Marker and ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Eisenia fetida Tissue sections (Ventral nerve cord)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Specification
Ventral nerve cord
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative
Formaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

MR. Carl Hobbs

Verified customer

Submitted Oct 24 2021

Western blot abreview for Anti-Histone H3 antibody - Nuclear Marker and ChIP Grade

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (Cell line)
Gel Running Conditions
Reduced Denaturing (12%)
Loading amount
58 µg
Specification
Cell line
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Oct 15 2021

Question

I had a few questions regarding ab1791. Does the Ab1791 antibody recognize all the variant forms of H3? Do you know the ratio of expression between the different variant forms in human cell lines?

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Abcam community

Verified customer

Asked on Oct 05 2012

Answer

Our antibody ab1791 will recognize total Histone H3 in your samples, so it will recognize the modified forms of Histone H3. Unfortunately I am unaware of the ratio of expression between the different variant forms in human cell lines.

Read More

Abcam Scientific Support

Answered on Oct 05 2012

Question

I am looking to conduct and IF assay using ab1791, with human samples. However, the host of the antibody cannot be rabbit. Is ab1791 a suitable candidate for the antibody? If not, could you please recommend another antibody?

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Abcam community

Verified customer

Asked on Mar 20 2012

Answer

I am happy to let you know that we have non-rabbit anti-human Histone H3 antibodies available, which also qualify for our testing discount programme (either because of their potential suitability for IF or because we do not have an image yet).

These are:

ab10799: mouse monoclonal, suitable for IHC, so might work for ICC/IF

https://www.abcam.com/Histone-H3-antibody-mAbcam-10799-ChIP-Grade-ab10799.html (or use the following: https://www.abcam.com/Histone-H3-antibody-mAbcam-10799-ChIP-Grade-ab10799.html).

ab128012: sheep polyclonal, suitable for ICC/IF, no image yet

https://www.abcam.com/Histone-H3-antibody-ab128012.html (or use the following: https://www.abcam.com/Histone-H3-antibody-ab128012.html).

Also, I could recommend our favourite rabbit polyclonal ab1791. I assume the customer is performing a costaining with another rabbit antibody and thus wants to avoid a rabbit anti-Histone H3 antibody. However, I am pleased to let you know that with our Easy Link kits this problem can be solved: You can easily label any primary antibody with a conjugate of your choice and eliminate the need for secondary antibodies. Please find more information under https://www.abcam.com/Easylink.

Read More

Abcam Scientific Support

Answered on Mar 20 2012

1-10 of 281 Abreviews or Q&A

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