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Rabbit Recombinant Monoclonal H3 mutated K36M antibody. Suitable for ChIC/CUT&RUN-seq, IP, WB, IHC-P, I-ELISA, ICC/IF, Flow Cyt (Intra) and reacts with Transfected cell line - Human, Human, Transfected cell lysate - Human, Synthetic peptide - Human samples.

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Images

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.05% 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.
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Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
ChIC/CUT&RUN-seqIPWBIHC-PI-ELISAICC/IFFlow Cyt (Intra)ChIP
Human
Expected
Expected
Expected
Tested
Expected
Tested
Tested
Not recommended
Mouse
Not recommended
Not recommended
Not recommended
Not recommended
Not recommended
Not recommended
Not recommended
Not recommended
Rat
Not recommended
Not recommended
Not recommended
Not recommended
Not recommended
Not recommended
Not recommended
Not recommended
Synthetic peptide - Human
Not recommended
Not recommended
Not recommended
Not recommended
Tested
Not recommended
Not recommended
Not recommended
Transfected cell line - Human
Tested
Not recommended
Not recommended
Not recommended
Not recommended
Not recommended
Not recommended
Not recommended
Transfected cell lysate - Human
Not recommended
Tested
Tested
Not recommended
Not recommended
Not recommended
Not recommended
Not recommended

Tested
Tested

Species

Transfected cell line - Human

Dilution info

3 µg

Notes

-

Expected
Expected

Species

Human

Dilution info

Use at an assay dependent concentration.

Notes

-

Not recommended
Not recommended

Species

Transfected cell lysate - Human, Mouse, Rat, Synthetic peptide - Human

Dilution info

-

Notes

-

Tested
Tested

Species

Transfected cell lysate - Human

Dilution info

1/30

Notes

-

Expected
Expected

Species

Human

Dilution info

Use at an assay dependent concentration.

Notes

-

Not recommended
Not recommended

Species

Transfected cell line - Human, Mouse, Rat, Synthetic peptide - Human

Dilution info

-

Notes

-

Tested
Tested

Species

Transfected cell lysate - Human

Dilution info

1/1000

Notes

-

Expected
Expected

Species

Human

Dilution info

Use at an assay dependent concentration.

Notes

-

Not recommended
Not recommended

Species

Transfected cell line - Human, Mouse, Rat, Synthetic peptide - Human

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

1/4000

Notes

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0)

Not recommended
Not recommended

Species

Mouse

Dilution info

-

Notes

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0)

Species

Rat

Dilution info

-

Notes

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0)

Species

Transfected cell line - Human

Dilution info

-

Notes

-

Species

Transfected cell lysate - Human

Dilution info

-

Notes

-

Species

Synthetic peptide - Human

Dilution info

-

Notes

-

Tested
Tested

Species

Synthetic peptide - Human

Dilution info

1 µg/mL

Notes

-

Expected
Expected

Species

Human

Dilution info

Use at an assay dependent concentration.

Notes

-

Not recommended
Not recommended

Species

Transfected cell line - Human, Transfected cell lysate - Human, Mouse, Rat

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

1/50

Notes

-

Not recommended
Not recommended

Species

Transfected cell line - Human, Transfected cell lysate - Human, Mouse, Rat, Synthetic peptide - Human

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

1/500

Notes

-

Not recommended
Not recommended

Species

Transfected cell line - Human, Transfected cell lysate - Human, Mouse, Rat, Synthetic peptide - Human

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Human, Transfected cell line - Human, Transfected cell lysate - Human, Mouse, Rat, Synthetic peptide - Human

Dilution info

-

Notes

-

Target data

Function

Variant histone H3 which replaces conventional H3 in a wide range of nucleosomes in active genes. Constitutes the predominant form of histone H3 in non-dividing cells and is incorporated into chromatin independently of DNA synthesis. Deposited at sites of nucleosomal displacement throughout transcribed genes, suggesting that it represents an epigenetic imprint of transcriptionally active chromatin. 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

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Rabbit Recombinant Monoclonal H3 mutated K36M antibody. Suitable for ChIC/CUT&RUN-seq, IP, WB, IHC-P, I-ELISA, ICC/IF, Flow Cyt (Intra) and reacts with Transfected cell line - Human, Human, Transfected cell lysate - Human, Synthetic peptide - Human samples.

Alternative names

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

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

EPR23614-91

Purification technique

Affinity purification Protein A

Concentration
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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.

9 product images

  • Immunoprecipitation - Anti-Histone H3 (mutated K36M) antibody [EPR23614-91] (ab256384), expandable thumbnail

    Immunoprecipitation - Anti-Histone H3 (mutated K36M) antibody [EPR23614-91] (ab256384)

    Histone H3.3 (mutated K36 M) was immunoprecipitated from 0.35 mg HEK-293 transfected with Histone H3.3 K36M (mutate) expression vector containing a myc-His-tag®, whole cell lysate with ab256384 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab256384 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

    Lane 1: HEK-293 transfected with Histone H3.3 K36M (mutate) expression vector containing a myc-His-tag®, whole cell lysate 10 ug

    Lane 2: ab256384 IP in HEK-293 transfected with Histone H3.3 K36M (mutate) expression vector containing a myc-His-tag®, whole cell lysate

    Lane 3:Rabbit monoclonal IgG (ab172730) instead of ab256384 in HEK-293 transfected with Histone H3.3 K36M (mutate) expression vector containing a myc-His-tag®, whole cell lysate

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 10 seconds

    All lanes: Immunoprecipitation - Anti-Histone H3 (mutated K36M) antibody [EPR23614-91] (AB256384)

    Predicted band size: 15 kDa

  • Western blot - Anti-Histone H3 (mutated K36M) antibody [EPR23614-91] (ab256384), expandable thumbnail

    Western blot - Anti-Histone H3 (mutated K36M) antibody [EPR23614-91] (ab256384)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST

    Exposure time: 26 seconds

    All lanes: Western blot - Anti-Histone H3 (mutated K36M) antibody [EPR23614-91] (AB256384) at 1/1000 dilution

    Lane 1: HEK-293 transfected with Histone H3.3 (WT) expression vector containing a myc-His-tag®, whole cell lysate at 10 µg

    Lane 2: HEK-293 transfected with Histone H3.3 K36M (mutate) expression vector containing a myc-His-tag®, whole cell lysate at 10 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051) at 1/100000 dilution

    Predicted band size: 15 kDa

    Observed band size: 20 kDa

  • Flow Cytometry (Intracellular) - Anti-Histone H3 (mutated K36M) antibody [EPR23614-91] (ab256384), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-Histone H3 (mutated K36M) antibody [EPR23614-91] (ab256384)

    Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HEK-293T transfected with myc tagged Histone H3 construct (Left) or myc tagged Histone H3 K36M construct (Right) cells labelling Histone H3 (mutated K36 M) with ab256384 at 1/500 compared with a isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 was used as the secondary antibody.

  • Indirect ELISA - Anti-Histone H3 (mutated K36M) antibody [EPR23614-91] (ab256384), expandable thumbnail

    Indirect ELISA - Anti-Histone H3 (mutated K36M) antibody [EPR23614-91] (ab256384)

    Indirect ELISA using ab256384 at varying antibody concentrations and antigen concentration at 100 ng/mL. An Alkaline Phosphatase-conjugated Goat Anti-Rabbit IgG (H+L) (1/2500) was used as the secondary antibody.

    Binding was seen for Histone H3 (mutated K36M) peptide. Binding to the following peptides was not seen:

    Histone H3 WT,
    Histone H3 (mono methyl K36),
    Histone H3 (di methyl K36),
    Histone H3 (tri methyl K36).

    This indicates the specificity of ab256384 for mutated K36M of Histone H3.

  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (mutated K36M) antibody [EPR23614-91] (ab256384), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (mutated K36M) antibody [EPR23614-91] (ab256384)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HEK-293 cells labelling Histone H3 (mutated K36 M) with ab256384 at 1/50 dilution, followed by a secondary antibody at 1/1000 dilution (Green). Confocal image showing nuclear staining in HEK-293 cell line transfected with myc-tagged Histone H3 K36M expression vector. 2233S Myc-Tag (9B11) Mouse mAb (Alexa Fluor® 647 Conjugate) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (mutated K36M) antibody [EPR23614-91] (ab256384), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (mutated K36M) antibody [EPR23614-91] (ab256384)

    Immunohistochemical analysis of paraffin-embedded human giant cell tumor of bone tissue labeling Histone H3 (mutated K36 M) with ab256384 at 1/4000 dilution followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Negative control: No staining in human giant cell tumor of bone (PMID: 29757500). Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

    Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0)

  • Western blot - Anti-Histone H3 (mutated K36M) antibody [EPR23614-91] (ab256384), expandable thumbnail

    Western blot - Anti-Histone H3 (mutated K36M) antibody [EPR23614-91] (ab256384)

    Blocking/Diluting buffer and concentration: 5% NFDM/TBST

    All lanes: Western blot - Anti-Histone H3 (mutated K36M) antibody [EPR23614-91] (AB256384) at 1/1000 dilution

    Lane 1: HEK-293 transfected with Histone H3.1 (WT) expression vector containing a myc-His-tag®, whole cell lysate at 20 µg

    Lane 2: HEK-293 transfected with Histone H3.1 K36M (mutant) expression vector containing a myc-His-tag®, whole cell lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051) at 1/100000 dilution

    Predicted band size: 15 kDa

    Observed band size: 19 kDa

    Exposure time: 62s

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (mutated K36M) antibody [EPR23614-91] (ab256384), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (mutated K36M) antibody [EPR23614-91] (ab256384)

    Immunohistochemical analysis of paraffin-embedded human chondroblastoma tissue labeling Histone H3 (mutated K36 M) with ab256384 at 1/4000 dilution followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Nuclear staining in human chondroblastoma (PMID: 29757500). Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

    Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0)

  • ChIC/CUT&RUN sequencing - Anti-Histone H3 (mutated K36M) antibody [EPR23614-91] (ab256384), expandable thumbnail

    ChIC/CUT&RUN sequencing - Anti-Histone H3 (mutated K36M) antibody [EPR23614-91] (ab256384)

    ChIC/CUT&RUN was performed using a pAG-MNAse at a final concentration of 700 ng/mL, 2.5 x 10^5 293T cells transfected with a H3.3K36M (Mutant) or H3.3 (WT) plasmid and 3µg of ab256384 [EPR23614-91]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown.

    Additional screenshots of mapped reads can be found in the Protocol booklet in the Support and downloads section.

    The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.

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'.

For licensing inquiries, please contact partnerships@abcam.com

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