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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.
IgG
Rabbit
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Monoclonal
ChIP | Dot | WB | ICC/IF | Flow 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 |
Species | Dilution info | Notes |
---|---|---|
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. |
Species | Dilution info | Notes |
---|---|---|
Species Synthetic peptide, Transfected cell lysate, Transfected cell line | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Synthetic peptide | Dilution info 1/1000 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Transfected cell lysate, Transfected cell line | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Transfected cell lysate | Dilution info 1/1000 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Synthetic peptide, Transfected cell line | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/100 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Synthetic peptide, Transfected cell lysate, Transfected cell line | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Transfected cell line | Dilution info 1/50 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Synthetic peptide, Transfected cell lysate | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
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. |
Species | Dilution info | Notes |
---|---|---|
Species Synthetic peptide, Transfected cell lysate, Transfected cell line | Dilution info - | Notes - |
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.
Histone H3.3, H3F3A, H3-3A, H3.3A, H3F3, PP781, H3-3B, H3.3B, H3F3B
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.
Histone H3.3, H3F3A, H3-3A, H3.3A, H3F3, PP781, H3-3B, H3.3B, H3F3B
IgG
Rabbit
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Monoclonal
The exact immunogen used to generate this antibody is proprietary information.
EPR23581-39
Affinity purification Protein A
Blue Ice
1-2 weeks
+4°C
-20°C
Upon delivery aliquot
Avoid freeze / thaw cycle
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:
For more information, read more on recombinant antibodies.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our 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.
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.
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
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
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.
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.
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).
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 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.
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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