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

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Images

Western blot - Anti-Histone H3.3 (mutated K36M) antibody [EPR23526-89] (AB254400), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3.3 (mutated K36M) antibody [EPR23526-89] (AB254400), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3.3 (mutated K36M) antibody [EPR23526-89] (AB254400), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3.3 (mutated K36M) antibody [EPR23526-89] (AB254400), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-Histone H3.3 (mutated K36M) antibody [EPR23526-89] (AB254400), expandable thumbnail

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% 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
ICC/IFIPChIPDotWBIHC-PI-ELISAFlow Cyt (Intra)
Human
Expected
Not recommended
Not recommended
Expected
Expected
Tested
Tested
Expected
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
Not recommended
Not recommended
Not recommended
Tested
Not recommended
Not recommended
Not recommended
Not recommended
Transfected cell line
Tested
Not recommended
Not recommended
Not recommended
Tested
Not recommended
Not recommended
Tested

Tested
Tested

Species

Transfected cell line

Dilution info

1/1000

Notes

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

Expected
Expected

Species

Human

Dilution info

Use at an assay dependent concentration.

Notes

-

Not recommended
Not recommended

Species

Synthetic peptide, Mouse, Rat

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

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

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

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

Dilution info

-

Notes

-

Tested
Tested

Species

Synthetic peptide

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, Mouse, Rat

Dilution info

-

Notes

-

Tested
Tested

Species

Transfected cell line

Dilution info

1/1000

Notes

-

Expected
Expected

Species

Human

Dilution info

Use at an assay dependent concentration.

Notes

-

Not recommended
Not recommended

Species

Synthetic peptide, Mouse, Rat

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

1/1000

Notes

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

Not recommended
Not recommended

Species

Mouse

Dilution info

-

Notes

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

Species

Rat

Dilution info

-

Notes

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

Species

Transfected cell line

Dilution info

-

Notes

-

Species

Synthetic peptide

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

0.0065 g/mL

Notes

-

Not recommended
Not recommended

Species

Transfected cell line, Synthetic peptide, Mouse, Rat

Dilution info

-

Notes

-

Tested
Tested

Species

Transfected cell line

Dilution info

1/500

Notes

-

Expected
Expected

Species

Human

Dilution info

Use at an assay dependent concentration.

Notes

-

Not recommended
Not recommended

Species

Synthetic peptide, Mouse, Rat

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

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

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

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

EPR23526-89

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.

7 product images

  • Western blot - Anti-Histone H3.3 (mutated K36M) antibody [EPR23526-89] (ab254400), expandable thumbnail

    Western blot - Anti-Histone H3.3 (mutated K36M) antibody [EPR23526-89] (ab254400)

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

    Exposure time: 92 seconds

    All lanes: Western blot - Anti-Histone H3.3 (mutated K36M) antibody [EPR23526-89] (ab254400) at 1/1000 dilution

    Lane 1: HEK-293 transfected with Histone H3.3 H3K36M expression vector containing a myc-His-tag®, whole cell lysate at 10 µg

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

    Secondary

    All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Predicted band size: 15 kDa

    Observed band size: 20 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3.3 (mutated K36M) antibody [EPR23526-89] (ab254400), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3.3 (mutated K36M) antibody [EPR23526-89] (ab254400)

    Immunohistochemical analysis of paraffin-embedded Human chondroblastoma tissue labelling Histone H3 (mutated K36M) with ab254400 at 1/1000 (0.548 ug/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Counterstained with Hematoxylin.

    Nuclear staining in human chondroblastoma (PMID: 29757500).

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

    Heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0)

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3.3 (mutated K36M) antibody [EPR23526-89] (ab254400), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3.3 (mutated K36M) antibody [EPR23526-89] (ab254400)

    Immunohistochemical analysis of paraffin-embedded Human gaint cell tumor of bone tissue labelling Histone H3 (mutated K36M) with ab254400 at 1/1000 (0.548 ug/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Counterstained with Hematoxylin.

    Negative control: No staining in human gaint cell tumor of bone (PMID: 29757500).

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

    Heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0)

  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3.3 (mutated K36M) antibody [EPR23526-89] (ab254400), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-Histone H3.3 (mutated K36M) antibody [EPR23526-89] (ab254400)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T cells labelling Histone H3 (mutated K36M) with ab254400 at 1/1000 (0.548 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 (2 ug/ml) dilution (Green). Myc-Tag Mouse mAb (Alexa Fluor® 647) was used to counterstain tubulin at 1/200 (2.5 ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

    Confocal image showing nuclear staining in 293T cells transfected with myc-tagged H3K36M expression vector is observed.

    Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 (2 ug/ml) dilution.

  • Flow Cytometry (Intracellular) - Anti-Histone H3.3 (mutated K36M) antibody [EPR23526-89] (ab254400), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-Histone H3.3 (mutated K36M) antibody [EPR23526-89] (ab254400)

    Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized 293T transfected with myc tagged Histone H3.3 construct (Left) or myc tagged Histone H3.3 H3K36M construct (Right) cells labelling Histone H3 (mutated K36M) with ab254400 at 1/500 dilution (0.1ug). A Goat anti rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody.

  • Dot Blot - Anti-Histone H3.3 (mutated K36M) antibody [EPR23526-89] (ab254400), expandable thumbnail

    Dot Blot - Anti-Histone H3.3 (mutated K36M) antibody [EPR23526-89] (ab254400)

    Dot blot analysis of Histone H3.3 (mutated K36M) using ab254400 at 1/1000 (0.548 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100,000 dilution.

    Lane 1: Histone H3.3 (mutated K36M)peptide (aa28-40)

    Lane 2: Histone H3.3 (mutated K36M) peptide (aa29-42)

    Lane 3: Histone H3.3 (WT) peptide (aa28-42)

    Exposure time: 3 minutes

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

  • Indirect ELISA - Anti-Histone H3.3 (mutated K36M) antibody [EPR23526-89] (ab254400), expandable thumbnail

    Indirect ELISA - Anti-Histone H3.3 (mutated K36M) antibody [EPR23526-89] (ab254400)

    ELISA using ab254400 at varying antibody concentrations and antigen concentration at 100 ng/ml. An Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) (1/2500) was used as the secondary antibody.

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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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