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Anti-Histone H3 antibody (Tri Methyl-K27) [EPR18607] ab192985 is a rabbit monoclonal antibody that is used in Histone H3 western blotting, IHC and immunofluorescence. Suitable for human, mouse and rat samples.

- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation
- Antibody clone EPR18607 is cited in over 90 publications


Images

ChIP - Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade (AB192985), expandable thumbnail
  • ChIP-sequencing - Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade (AB192985), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade (AB192985), expandable thumbnail
  • Western blot - Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade (AB192985), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade (AB192985), expandable thumbnail

Publications

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
ChIPELISAWBPepArrICC/IFChIP-seqIHC-P
Human
Tested
Expected
Tested
Tested
Tested
Tested
Tested
Mouse
Expected
Tested
Tested
Expected
Expected
Expected
Tested
Rat
Expected
Expected
Expected
Expected
Expected
Expected
Tested

Tested
Tested

Species

Human

Dilution info

2 µg chromatin for 25.00000 µg chromatin

Notes

Use Myo-D ChIP primer pair ab269261 as positive control.

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

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Mouse

Dilution info

0.25 µg/mL

Notes

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

Expected
Expected

Species

Human, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Mouse

Dilution info

1/1000

Notes

We suggest to use 2% BSA as blocking and antibody dilution buffer. To get stronger band, 1%SDS Hot lysis method is also recommended.

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

Species

Human

Dilution info

1/1000

Notes

We suggest to use 2% BSA as blocking and antibody dilution buffer. To get stronger band, 1%SDS Hot lysis method is also recommended.

Expected
Expected

Species

Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Human

Dilution info

0.1 µg/mL

Notes

-

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Human

Dilution info

1/1000

Notes

-

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Human

Dilution info

4 µg/cells for 7.00000 µg/cells

Notes

-

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Mouse

Dilution info

1/500

Notes

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

Species

Rat

Dilution info

1/500

Notes

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

Species

Human

Dilution info

1/500

Notes

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

Associated Products

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Target data

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.

Alternative names

Recommended products

Anti-Histone H3 antibody (Tri Methyl-K27) [EPR18607] ab192985 is a rabbit monoclonal antibody that is used in Histone H3 western blotting, IHC and immunofluorescence. Suitable for human, mouse and rat samples.

- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation
- Antibody clone EPR18607 is cited in over 90 publications

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number

EPR18607

Purification technique

Affinity purification Protein A

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

Notes

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

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 batch production

For more information, read more on recombinant antibodies.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

14 product images

  • ChIP - Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade (ab192985), expandable thumbnail

    ChIP - Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade (ab192985)

    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 ab192985 (blue), and 20 μl of Anti rabbit IgG sepharose beads. 2 μg of rabbit normal IgG was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).

  • ChIP-sequencing - Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade (ab192985), expandable thumbnail

    ChIP-sequencing - Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade (ab192985)

    Chromatin was prepared from HeLa cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 107 HeLa cells and 4 μg of Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade (ab192985). ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads.

    Additional screenshots of mapped reads can be downloaded here.

  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade (ab192985), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade (ab192985)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling Histone H3 (tri methyl K27) with ab192985 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on HeLa cell line. The nuclear counterstain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin mouse MAb (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) secondary antibody at 1/1000 dilution (red).

    The negative controls are as follows:-
    -ve control 1: ab192985 at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) secondary antibody at 1/1000 dilution.
    -ve control 2: Anti-alpha Tubulin mouse MAb (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution.

  • Western blot - Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade (ab192985), expandable thumbnail

    Western blot - Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade (ab192985)

    Blocking/Diluting buffer: 5% NFDM/TBST

    For this product, we recommend 1% SDS hot lysis method.

    For Lysate preparation protocol, please refer to the protocol section of the website and/or here.

    All lanes: Western blot - Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade (ab192985) at 1/1000 dilution

    Lane 1: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate prepared using 1% SDS hot lysis method at 20 µg

    Lane 2: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate prepared using RIPA lysis method at 20 µg

    Secondary

    All lanes: Goat Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/5000 dilution

    Predicted band size: 15 kDa

    Observed band size: 15 kDa

    Exposure time: 3min

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade (ab192985), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade (ab192985)

    Immunohistochemical analysis of paraffin-embedded human colon tissue labeling Histone H3 (tri methyl K27) with ab192985 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution. Nuclear staining on human colon tissue 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) (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.

  • Western blot - Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade (ab192985), expandable thumbnail

    Western blot - Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade (ab192985)

    We recommend to use 2% BSA as blocking and antibody dilution buffer.

    All lanes: Western blot - Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade (ab192985) at 1/1000 dilution

    All lanes: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates at 15 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051)

    Predicted band size: 15 kDa

    Observed band size: 15 kDa

    Exposure time: 40s

  • Western blot - Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade (ab192985), expandable thumbnail

    Western blot - Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade (ab192985)

    Blocking/Dilution buffer: 5% BSA/TBST

    All lanes: Western blot - Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade (ab192985) at 1/1000 dilution

    All lanes: NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate at 10 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/50000 dilution

    Predicted band size: 15 kDa, 45 kDa

    Observed band size: 15 kDa, 30 kDa

    Exposure time: 1s

  • Western blot - Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade (ab192985), expandable thumbnail

    Western blot - Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade (ab192985)

    The antibody is blocked by tri methyl K27 peptide (lane 10) and slightly by di methyl K27 peptide (lane 9, there is 14% cross reactivity with di methyl K27 as determined by ELISA).

    All lanes: Western blot - Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade (ab192985) at 1/1000 dilution

    All lanes: HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 10 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

    Developed using the ECL technique.

    Predicted band size: 15 kDa

    Exposure time: 5s

  • Western blot - Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade (ab192985), expandable thumbnail

    Western blot - Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade (ab192985)

    All lanes: Western blot - Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade (ab192985) at 1/1000 dilution

    Lane 1: HeLa (Human epithelial cell line from cervix adenocarcinoma) at 10 µg

    Lane 2: EED-/- mouse whole cell lysate at 10 µg

    Lane 3: Wild type mouse ES whole cell lysate at 10 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/50000 dilution

    Developed using the ECL technique.

    Predicted band size: 15 kDa

    Exposure time: 8min

  • Peptide Array - Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade (ab192985), expandable thumbnail

    Peptide Array - Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade (ab192985)

    Peptide array analysis was performed using ab192985 at a concentration of 0.1 µg/ml, followed by Goat Anti-Rabbit IgG, (H+L), Fluo 647nm conjugated secondary antibody at a 1/50,000 dilution.

    ab192985 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, is available under the Support & downloads section.

  • ELISA - Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade (ab192985), expandable thumbnail

    ELISA - Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade (ab192985)

    ELISA analysis was performed on 1 μg/ml of antigen using ab192985 at a concentration range of 0-0.25 μg/ml, followed by Alkaline Phosphatase-conjugates AffiniPure Goat anti-rabbit IgG (H&L) secondary antibody at a 1/2,500 dilution.

    All batches of ab192985 are tested in ELISA against peptides to different Histone H3 modifications. Results show strong binding to Histone H3 - tri methyl K27 immunizing peptide, indicating that this antibody specifically recognizes the Histone H3 - tri methyl K27 modification. Weak binding (14%) was also detected against H3 - di methyl K27 modification.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade (ab192985), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade (ab192985)

    Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling Histone H3 (tri methyl K27) with ab192985 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution. Nuclear staining on mouse colon tissue is observed. Counterstained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (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 (tri methyl K27) antibody [EPR18607] - ChIP Grade (ab192985), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade (ab192985)

    Immunohistochemical analysis of paraffin-embedded rat kidney tissue labeling Histone H3 (tri methyl K27) with ab192985 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution. Nuclear staining on rat kidney tissue is observed. Counterstained with hematoxylin.

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

  • ChIP-sequencing - Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade (ab192985), expandable thumbnail
    This image is courtesy of a customer review submitted by Andrew Perez

    ChIP-sequencing - Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade (ab192985)

    Chromatin was prepared from human Nuclear cell lysate K562 cells.
    Cells were fixed with 1% formaldehyde for 10 minutes. Cells were incubated with the primary antibody with PBS + 0.1% Tween-20 for 1 hour at 25°C

Downloads

Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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