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.
IgG
Rabbit
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Monoclonal
ICC/IF | IP | ChIP | Dot | WB | IHC-P | I-ELISA | Flow 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 |
Species | Dilution info | Notes |
---|---|---|
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. |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Synthetic peptide, Mouse, Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human, Transfected cell line, Synthetic peptide, Mouse, Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human, Transfected cell line, Synthetic peptide, Mouse, Rat | 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 line, Mouse, Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Transfected cell line | 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, Mouse, Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
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. |
Species | Dilution info | Notes |
---|---|---|
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 - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 0.0065 g/mL | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Transfected cell line, Synthetic peptide, Mouse, Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Transfected cell line | Dilution info 1/500 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Synthetic peptide, Mouse, Rat | 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.
H3.3A, H3F3, H3F3A, PP781, H3-3B, H3.3B, H3F3B, H3-3A, Histone H3.3
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.
IgG
Rabbit
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Monoclonal
EPR23526-89
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.
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
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
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)
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)
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 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 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
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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