Hello. We're improving abcam.com and we'd welcome your feedback.

Hello. We're improving abcam.com and we'd welcome your feedback.

Infomation icon

We haven't added this to the BETA yet

New BETA website

New BETA website

Hello. We're improving abcam.com and we'd welcome your feedback.

Take a look at our BETA site and see what we’ve done so far.

Switch on our new BETA site

Now available

Search and browse selected products

  • A selection of primary antibodies

Purchase these through your usual distributor

In the coming months

  • Additional product types
  • Supporting content
  • Sign in to your account
  • Purchase online
RecombinantRabMAb

Recombinant Anti-Histone H3 (di methyl K9) antibody [EP16990] - ChIP Grade (ab176882)

Reviews (4) Submit a question References (6)
ChIP - Anti-Histone H3 (di methyl K9) antibody [EP16990] - ChIP Grade (ab176882)
  • Peptide Array - Anti-Histone H3 (di methyl K9) antibody [EP16990] - ChIP Grade (ab176882)
  • Western blot - Anti-Histone H3 (di methyl K9) antibody [EP16990] - ChIP Grade (ab176882)
  • Western blot - Anti-Histone H3 (di methyl K9) antibody [EP16990] - ChIP Grade (ab176882)
  • Western blot - Anti-Histone H3 (di methyl K9) antibody [EP16990] - ChIP Grade (ab176882)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (di methyl K9) antibody [EP16990] - ChIP Grade (ab176882)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (di methyl K9) antibody [EP16990] - ChIP Grade (ab176882)
  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K9) antibody [EP16990] - ChIP Grade (ab176882)
  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K9) antibody [EP16990] - ChIP Grade (ab176882)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP16990] to Histone H3 (di methyl K9) - ChIP Grade
  • Suitable for: PepArr, ChIP, WB, ICC/IF, IHC-P
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-Histone H3 (di methyl K9) antibody [EP16990] - ChIP Grade
    See all Histone H3 primary antibodies

  • Description

    Rabbit monoclonal [EP16990] to Histone H3 (di methyl K9) - ChIP Grade

  • Host species

    Rabbit

  • Tested applications

    Suitable for: PepArr, ChIP, WB, ICC/IF, IHC-Pmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: NIH/3T3, HeLa and PC-12 whole cell lysates. IHC-P: Mouse spleen and rat kidney tissues. ICC/IF: HeLa cells. ChIP: Chromatin prepared from HeLa cells.

  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    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. Avoid freeze / thaw cycle.

  • Storage buffer

    Preservative: 0.01% Sodium azide
    Constituents: PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EP16990

  • Isotype

    IgG

  • Research areas

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab176882 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
PepArr
Use at an assay dependent concentration.
ChIP
Use 2 µg for 25 µg of chromatin.
WB (3)
1/1000. Detects a band of approximately 15 kDa (predicted molecular weight: 15 kDa).
ICC/IF
1/500.
IHC-P (1)
1/50. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Notes
PepArr
Use at an assay dependent concentration.
ChIP
Use 2 µg for 25 µg of chromatin.
WB
1/1000. Detects a band of approximately 15 kDa (predicted molecular weight: 15 kDa).
ICC/IF
1/500.
IHC-P
1/50. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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.
  • Information by UniProt

  • Database links

    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

  • ChIP - Anti-Histone H3 (di methyl K9) antibody [EP16990] - ChIP Grade (ab176882)
    ChIP - Anti-Histone H3 (di methyl K9) antibody [EP16990] - ChIP Grade (ab176882)

    Chromatin was prepared from HeLa cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 2µg of ab176882 (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 for active and inactive loci, Sybr green approach for heterochromatic loci). Primers and probes are located in the first kb of the transcribed region.

  • Peptide Array - Anti-Histone H3 (di methyl K9) antibody [EP16990] - ChIP Grade (ab176882)
    Peptide Array - Anti-Histone H3 (di methyl K9) antibody [EP16990] - ChIP Grade (ab176882)

    ab176882 was tested in Peptide array against 501 different modified and unmodified histone peptides; each peptide is printed on the array at six concentrations (each in triplicate).
     
    Circle area represents affinity between the antibody and a peptide: all antigen-containing peptides are displayed as red circles, all other peptides as blue circles. The affinity is calculated as area under curve when antibody binding values are plotted against the corresponding peptide concentration. Each circle area is normalized to the peptide with the strongest affinity.
     
    The complete dataset, including full list of all peptides and information on the position of each peptide in the diagram, can be downloaded here.

  • Western blot - Anti-Histone H3 (di methyl K9) antibody [EP16990] - ChIP Grade (ab176882)
    Western blot - Anti-Histone H3 (di methyl K9) antibody [EP16990] - ChIP Grade (ab176882)
    Anti-Histone H3 (di methyl K9) antibody [EP16990] - ChIP Grade (ab176882) at 1/1000 dilution + NIH/3T3 (Mouse embryonic fibroblast cell line) cell lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 15 kDa
    Observed band size: 15 kDa


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-Histone H3 (di methyl K9) antibody [EP16990] - ChIP Grade (ab176882)
    Western blot - Anti-Histone H3 (di methyl K9) antibody [EP16990] - ChIP Grade (ab176882)
    Anti-Histone H3 (di methyl K9) antibody [EP16990] - ChIP Grade (ab176882) at 1/100000 dilution + HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 15 kDa
    Observed band size: 15 kDa


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-Histone H3 (di methyl K9) antibody [EP16990] - ChIP Grade (ab176882)
    Western blot - Anti-Histone H3 (di methyl K9) antibody [EP16990] - ChIP Grade (ab176882)
    Anti-Histone H3 (di methyl K9) antibody [EP16990] - ChIP Grade (ab176882) at 1/1000 dilution + PC-12 (Rat adrenal gland pheochromocytoma cell line) cell lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/100000 dilution

    Predicted band size: 15 kDa
    Observed band size: 15 kDa


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (di methyl K9) antibody [EP16990] - ChIP Grade (ab176882)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (di methyl K9) antibody [EP16990] - ChIP Grade (ab176882)

    Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling Histone H3 (di methyl K9) with ab176882 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on mouse spleen is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (di methyl K9) antibody [EP16990] - ChIP Grade (ab176882)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (di methyl K9) antibody [EP16990] - ChIP Grade (ab176882)

    Immunohistochemical analysis of paraffin-embedded rat kidney tissue labeling Histone H3 (di methyl K9) with ab176882 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on rat kidney is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K9) antibody [EP16990] - ChIP Grade (ab176882)
    Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K9) antibody [EP16990] - ChIP Grade (ab176882)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Histone H3 (di methyl K9) with ab176882 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Confocal image showing nuclear staining on HeLa cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/500 dilution (red).

    The negative controls are as follows:
    -ve control 1: ab176882 at 1/500 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/500 dilution.
    -ve control 2: Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution.

  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K9) antibody [EP16990] - ChIP Grade (ab176882)
    Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K9) antibody [EP16990] - ChIP Grade (ab176882)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Histone H3 (di methyl K9) with ab176882 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Confocal image showing nuclear staining on HeLa cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/500 dilution (red).

    The negative controls are as follows:
    -ve control 1: ab176882 at 1/500 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/500 dilution.
    -ve control 2: Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution.

References (6)

Publishing research using ab176882? Please let us know so that we can cite the reference in this datasheet.

ab176882 has been referenced in 6 publications.

  • Claycombe-Larson KG  et al. Paternal high-fat diet and exercise regulate sperm miRNA and histone methylation to modify placental inflammation, nutrient transporter mRNA expression and fetal weight in a sex-dependent manner. J Nutr Biochem 81:108373 (2020). PubMed: 32422425
  • Glanzner WG  et al. Histone Lysine Demethylases KDM5B and KDM5C Modulate Genome Activation and Stability in Porcine Embryos. Front Cell Dev Biol 8:151 (2020). PubMed: 32211412
  • Viscarra JA  et al. Histone demethylase JMJD1C is phosphorylated by mTOR to activate de novo lipogenesis. Nat Commun 11:796 (2020). PubMed: 32034158
  • Stegen S  et al. Glutamine Metabolism Controls Chondrocyte Identity and Function. Dev Cell 53:530-544.e8 (2020). PubMed: 32470321
  • Grosse L  et al. Defined p16High Senescent Cell Types Are Indispensable for Mouse Healthspan. Cell Metab 32:87-99.e6 (2020). PubMed: 32485135
  • Zhang W  et al. Knockdown of KDM1A suppresses tumour migration and invasion by epigenetically regulating the TIMP1/MMP9 pathway in papillary thyroid cancer. J Cell Mol Med 23:4933-4944 (2019). PubMed: 31211500

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

1-4 of 4 Abreviews or Q&A

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Histone H3 (di methyl K9) antibody [EP16990] - ChIP Grade

 Good
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Pancreatic tumor)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris/EDTA pH9
Permeabilization
No
Specification
Pancreatic tumor
Blocking step
Serum as blocking agent for 20 minute(s) · Concentration: 2.5% · Temperature: 20°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.

Clara Lubeseder-Martellato

Verified customer

Submitted Jun 28 2021

Western blot abreview for Anti-Histone H3 (di methyl K9) antibody [EP16990] - ChIP Grade

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Mouse Cell lysate - other (Pancreatic tumors cells)
Gel Running Conditions
Reduced Denaturing (12.5)
Loading amount
15 µg
Specification
Pancreatic tumors cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°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.

Clara Lubeseder-Martellato

Verified customer

Submitted Jun 16 2021

Western blot abreview for Anti-Histone H3 (di methyl K9) antibody [EP16990] - ChIP Grade

 Average
Abreviews
abreview image
Application
Western blot
Sample
Human Purified protein (K-562)
Gel Running Conditions
Reduced Denaturing (18% gel)
Loading amount
0.5 µg
Specification
K-562
Blocking step
Odyssey Blocking Buffer (TBS) (LI-COR) 1:1 mixture of Licor blocking buffer and 1X PBS as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · 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.

Dr. Ragnhild Eskeland

Verified customer

Submitted Nov 14 2016

Western blot abreview for Anti-Histone H3 (di methyl K9) antibody [EP16990] - ChIP Grade

 Good
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - nuclear (Melanoma)
Gel Running Conditions
Reduced Denaturing (15%)
Loading amount
3000 cells
Specification
Melanoma
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°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 08 2016

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com