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Rabbit Recombinant Monoclonal H3 mutated G34W antibody. Suitable for ChIP, Dot, WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Human, Synthetic peptide, Transfected cell lysate, Transfected cell line samples.

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Images

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.
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Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
ChIPDotWBICC/IFFlow Cyt (Intra)IHC-P
Human
Tested
Expected
Expected
Tested
Expected
Tested
Synthetic peptide
Not recommended
Tested
Not recommended
Not recommended
Not recommended
Not recommended
Transfected cell line
Not recommended
Not recommended
Not recommended
Not recommended
Tested
Not recommended
Transfected cell lysate
Not recommended
Not recommended
Tested
Not recommended
Not recommended
Not recommended

Tested
Tested

Species

Human

Dilution info

2 µg for 25.00000 µg chromatin

Notes

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

Not recommended
Not recommended

Species

Synthetic peptide, Transfected cell lysate, Transfected cell line

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 lysate, Transfected cell line

Dilution info

-

Notes

-

Tested
Tested

Species

Transfected cell lysate

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, Transfected cell line

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

1/100

Notes

-

Not recommended
Not recommended

Species

Synthetic peptide, Transfected cell lysate, Transfected cell line

Dilution info

-

Notes

-

Tested
Tested

Species

Transfected cell line

Dilution info

1/50

Notes

-

Expected
Expected

Species

Human

Dilution info

Use at an assay dependent concentration.

Notes

-

Not recommended
Not recommended

Species

Synthetic peptide, Transfected cell lysate

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

1/250

Notes

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

Not recommended
Not recommended

Species

Synthetic peptide, Transfected cell lysate, Transfected cell line

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

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Rabbit Recombinant Monoclonal H3 mutated G34W antibody. Suitable for ChIP, Dot, WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Human, Synthetic peptide, Transfected cell lysate, Transfected cell line 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

EPR23581-39

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.

7 product images

  • ChIP - Anti-Histone H3.3 (mutated G34W) antibody [EPR23581-39] - ChIP Grade (ab272691), expandable thumbnail

    ChIP - Anti-Histone H3.3 (mutated G34W) antibody [EPR23581-39] - ChIP Grade (ab272691)

    Chromatin was prepared from HEK-293T transfected with myc-His tagged Histone H3.3 mutated G34W and Histone H3.3 WT cells according to the Abcam Dual-X-ChIP protocol. Cells were fixed with formaldehyde for 10min.
    The ChIP was performed with 25 μg of chromatin, 2 μg of ab272691 (red), or 2 μg of rabbit normal IgG ab172730 (gray) and 20 μl of Protein A/G sepharose beads. 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.

  • Western blot - Anti-Histone H3.3 (mutated G34W) antibody [EPR23581-39] - ChIP Grade (ab272691), expandable thumbnail

    Western blot - Anti-Histone H3.3 (mutated G34W) antibody [EPR23581-39] - ChIP Grade (ab272691)

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

    Exposure time: 3 minutes.

    All lanes: Western blot - Anti-Histone H3.3 (mutated G34W) antibody [EPR23581-39] - ChIP Grade (AB272691) at 1/1000 dilution

    Lane 1: HEK-293T (human embryonic kidney) transfected with Histone H3.3 G34W expression vector containing a myc-His-tag®, whole cell lysate at 40 µg

    Lane 2: HEK-293T transfected with Histone H3.3 (WT) expression vector containing a myc-His-tag®, whole cell lysate at 40 µg

    Secondary

    All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/50000 dilution

    Predicted band size: 15 kDa

    Observed band size: 20 kDa

  • Flow Cytometry (Intracellular) - Anti-Histone H3.3 (mutated G34W) antibody [EPR23581-39] - ChIP Grade (ab272691), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-Histone H3.3 (mutated G34W) antibody [EPR23581-39] - ChIP Grade (ab272691)

    Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed 90% methanol-permeabilized HEK-293T (Human embryonic kidney epithelial cell) transfected with myc tagged Histone H3.3 WT construct (Left panel) and myc-tagged Histone H3.3 G34W construct (Right panel) cells labelling Histone H3.3 (mutated G34 W) with ab272691 at 1/50 dilution (1μg). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3.3 (mutated G34W) antibody [EPR23581-39] - ChIP Grade (ab272691), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-Histone H3.3 (mutated G34W) antibody [EPR23581-39] - ChIP Grade (ab272691)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HEK-293T cells labelling Histone H3.3(mutated G34 W) with ab272691 at 1/100 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 2 μg/ml dilution (Green). Confocal image showing nuclear staining in HEK-293T cells transfected with Histone H3.3 G34W-Myc plasmid, while no staining in HEK-293T cells transfected with H3.3 WT -Myc plasmid. Myc-Tag Mouse mAb (Alexa Fluor® 647) was used to counterstain tubulin at 1/200 dilution (Red). The nuclear counterstain was DAPI (Blue).

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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3.3 (mutated G34W) antibody [EPR23581-39] - ChIP Grade (ab272691), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3.3 (mutated G34W) antibody [EPR23581-39] - ChIP Grade (ab272691)

    Immunohistochemical analysis of paraffin-embedded human giant cell tumor of bone tissue labeling Histone H3.3(mutated G34 W) with ab272691 at 1/250 dilution followed by ready to use Goat Anti-Rabbit IgG H&L (HRP). Nuclear staining in human giant cell tumor of bone (PMID: 29757500). Counterstained with Hematoxylin.

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

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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3.3 (mutated G34W) antibody [EPR23581-39] - ChIP Grade (ab272691), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3.3 (mutated G34W) antibody [EPR23581-39] - ChIP Grade (ab272691)

    Immunohistochemical analysis of paraffin-embedded human chondroblastoma tissue labeling Histone H3.3(mutated G34 W) with ab272691 at 1/250 dilution followed by ready to use Goat Anti-Rabbit IgG H&L (HRP).

    Negative control: No staining in human chondroblastoma (PMID: 29757500).

    Counterstained with Hematoxylin.

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

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

  • Dot Blot - Anti-Histone H3.3 (mutated G34W) antibody [EPR23581-39] - ChIP Grade (ab272691), expandable thumbnail

    Dot Blot - Anti-Histone H3.3 (mutated G34W) antibody [EPR23581-39] - ChIP Grade (ab272691)

    Dot blot analysis of Histone H3.3 (mutated G34 W) labeled with ab272691 at 1/1000 dilution.

    Lane 1: Histone H3.3 H3G34W peptide (aa28-37).
    Lane 2: Histone H3.3 H3G34W peptide (aa33-43).
    Lane 3: Histone H3.3 H3G34W peptide (aa28-43).
    Lane 4: Histone H3.3 WT peptide (aa28-43).

    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution was used as secondary antibody.

    Blocking and dilution buffer: 5% NFDM/TBST.

    Exposure time: 3 minutes.

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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'.

For licensing inquiries, please contact partnerships@abcam.com

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