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Rabbit Recombinant Monoclonal H3 mutated G34V antibody. Suitable for IP, ChIP, Dot, WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Transfected cell lysate, Human, Transfected cell line, Synthetic peptide, Transfected cell lysate - Human 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
IPChIPDotWBICC/IFFlow Cyt (Intra)IHC-P
Human
Expected
Expected
Expected
Expected
Expected
Expected
Tested
Mouse
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
Synthetic peptide
Not recommended
Not recommended
Tested
Not recommended
Not recommended
Not recommended
Not recommended
Transfected cell line
Not recommended
Tested
Not recommended
Not recommended
Tested
Tested
Not recommended
Transfected cell lysate
Tested
Not recommended
Not recommended
Not recommended
Not recommended
Not recommended
Not recommended
Transfected cell lysate - Human
Not recommended
Not recommended
Not recommended
Tested
Not recommended
Not recommended
Not recommended

Tested
Tested

Species

Transfected cell lysate

Dilution info

-

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

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

Dilution info

-

Notes

-

Tested
Tested

Species

Transfected cell line

Dilution info

2 µg for 25 µg chromatin

Notes

-

Expected
Expected

Species

Human

Dilution info

Use at an assay dependent concentration.

Notes

-

Not recommended
Not recommended

Species

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

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 lysate, Transfected cell line, Synthetic peptide, 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

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

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

Dilution info

-

Notes

-

Species

Transfected cell line

Dilution info

-

Notes

-

Species

Synthetic peptide

Dilution info

-

Notes

-

Species

Transfected cell lysate - 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

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Rabbit Recombinant Monoclonal H3 mutated G34V antibody. Suitable for IP, ChIP, Dot, WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Transfected cell lysate, Human, Transfected cell line, Synthetic peptide, Transfected cell lysate - 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

EPR23520-5

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.

8 product images

  • ChIP - Anti-Histone H3.3 (mutated G34V) antibody [EPR23520-5] - ChIP Grade (ab254401), expandable thumbnail

    ChIP - Anti-Histone H3.3 (mutated G34V) antibody [EPR23520-5] - ChIP Grade (ab254401)

    Chromatin was prepared from HEK-293T transfected with myc-His tagged Histone H3.3 mutated G34V and Histone H3.3 WT cells according to the Abcam Dual-X-ChIP protocol. Cells were fixed with then formaldehyde for 10min.
    The ChIP was performed with 25 μg of chromatin, 2 μg of ab254401 (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.

  • Immunoprecipitation - Anti-Histone H3.3 (mutated G34V) antibody [EPR23520-5] - ChIP Grade (ab254401), expandable thumbnail

    Immunoprecipitation - Anti-Histone H3.3 (mutated G34V) antibody [EPR23520-5] - ChIP Grade (ab254401)

    Histone H3.3(mutated G34 V) was immunoprecipitated from 0.35 mg HEK-293T (human embryonic kidney) transfected with Histone H3.3 G34V expression vector containing a myc-His-tag® whole cell lysate 10 μg with ab254401 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab254401 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

    Lane 1: HEK-293T transfected with Histone H3.3 G34V expression vector containing a myc-His-tag® whole cell lysate 10 μg

    Lane 2: ab254401 IP in HEK-293T transfected with Histone H3.3 G34V expression vector containing a myc-His-tag® whole cell lysate

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab254401 in HEK-293T transfected with Histone H3.3 G34V expression vector containing a myc-His-tag® whole cell lysate

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

    Exposure time: 32 seconds.

    All lanes: Immunoprecipitation - Anti-Histone H3.3 (mutated G34V) antibody [EPR23520-5] - ChIP Grade (AB254401)

    Predicted band size: 15 kDa

  • Western blot - Anti-Histone H3.3 (mutated G34V) antibody [EPR23520-5] - ChIP Grade (ab254401), expandable thumbnail

    Western blot - Anti-Histone H3.3 (mutated G34V) antibody [EPR23520-5] - ChIP Grade (ab254401)

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

    Exposure time: 10 seconds.

    All lanes: Western blot - Anti-Histone H3.3 (mutated G34V) antibody [EPR23520-5] - ChIP Grade (AB254401) at 1/1000 dilution

    Lane 1: HEK-293T (human embryonic kidney) transfected with Histone H3.3 G34V expression vector containing a myc-His-tag®, whole cell lysate at 20 µg

    Lane 2: HEK-293T transfected with Histone H3.3 (WT) expression vector containing a myc-His-tag®, whole cell lysate at 20 µg

    Secondary

    All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution

    Predicted band size: 15 kDa

    Observed band size: 20 kDa

  • Flow Cytometry (Intracellular) - Anti-Histone H3.3 (mutated G34V) antibody [EPR23520-5] - ChIP Grade (ab254401), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-Histone H3.3 (mutated G34V) antibody [EPR23520-5] - ChIP Grade (ab254401)

    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 H3G34V construct (Right panel) cells labelling Histone H3.3 (mutated G34 V) with ab254401 at 1/500 dilution (0.1μg). 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 G34V) antibody [EPR23520-5] - ChIP Grade (ab254401), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-Histone H3.3 (mutated G34V) antibody [EPR23520-5] - ChIP Grade (ab254401)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HEK-293T cells labelling Histone H3.3(mutated G34 V) with ab254401 at 1/1000 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 H3G34V-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 G34V) antibody [EPR23520-5] - ChIP Grade (ab254401), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3.3 (mutated G34V) antibody [EPR23520-5] - ChIP Grade (ab254401)

    Immunohistochemical analysis of paraffin-embedded human giant cell tumor of bone tissue labeling Histone H3.3(mutated G34 V) with ab254401 at 1/1000 dilution (0.542 μg/ml) dilution followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human giant cell tumor of bone. (PMID: 29241742). The section was incubated with ab254401 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3.3 (mutated G34V) antibody [EPR23520-5] - ChIP Grade (ab254401), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3.3 (mutated G34V) antibody [EPR23520-5] - ChIP Grade (ab254401)

    Immunohistochemical analysis of paraffin-embedded human chondroblastoma tissue labeling Histone H3.3(mutated G34 V) with ab254401 at 1/1000 dilution (0.542 μg/ml) dilution followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Negative control: No staining on human chondroblastoma (PMID: 29241742).

    The section was incubated with ab254401 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

  • Dot Blot - Anti-Histone H3.3 (mutated G34V) antibody [EPR23520-5] - ChIP Grade (ab254401), expandable thumbnail

    Dot Blot - Anti-Histone H3.3 (mutated G34V) antibody [EPR23520-5] - ChIP Grade (ab254401)

    Dot blot analysis of Histone H3.3 (mutated G34 V) labeled with ab254401 at 1/1000 dilution.

    Lane 1: Histone H3.3 H3G34V peptide (aa28-40).
    Lane 2: Histone H3.3 H3G34V peptide (aa26-38).
    Lane 3: Histone H3.3 WT peptide (aa26-40).

    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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