Rabbit Recombinant Monoclonal H3 mutated K36M antibody. Carrier free. Suitable for ICC/IF, Flow Cyt (Intra), Dot, WB, IHC-P, I-ELISA and reacts with Human, Transfected cell line samples.
IgG
Rabbit
pH: 7.2 - 7.4
Constituents: 100% PBS
Liquid
Monoclonal
ICC/IF | IP | ChIP | Flow Cyt (Intra) | Dot | WB | IHC-P | I-ELISA | |
---|---|---|---|---|---|---|---|---|
Human | Tested | Not recommended | Not recommended | Expected | Tested | Tested | Tested | Tested |
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 |
Transfected cell line | Not recommended | Not recommended | Not recommended | Tested | Not recommended | Not recommended | Not recommended | Not recommended |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info - | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Transfected cell line, Mouse, Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human, Transfected cell line, Mouse, Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human, Transfected cell line, Mouse, Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Transfected cell line | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Transfected cell line, Mouse, Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Transfected cell line, Mouse, Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info - | 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 | Dilution info | Notes |
---|---|---|
Species Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Transfected cell line, 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. Carrier free. Suitable for ICC/IF, Flow Cyt (Intra), Dot, WB, IHC-P, I-ELISA and reacts with Human, Transfected cell line samples.
IgG
Rabbit
pH: 7.2 - 7.4
Constituents: 100% PBS
Liquid
Monoclonal
Yes
EPR23526-89
Affinity purification Protein A
Blue Ice
+4°C
+4°C
ab277487 is the carrier-free version of Anti-Histone H3.3 (mutated K36M) antibody [EPR23526-89] ab254400.
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.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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.
This data was developed using Anti-Histone H3.3 (mutated K36M) antibody [EPR23526-89] ab254400, the same antibody clone in a different buffer formulation.
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] (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
This data was developed using Anti-Histone H3.3 (mutated K36M) antibody [EPR23526-89] ab254400, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Human chondroblastoma tissue labelling Histone H3 (mutated K36M) with Anti-Histone H3.3 (mutated K36M) antibody [EPR23526-89] 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)
This data was developed using Anti-Histone H3.3 (mutated K36M) antibody [EPR23526-89] ab254400, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Human gaint cell tumor of bone tissue labelling Histone H3 (mutated K36M) with Anti-Histone H3.3 (mutated K36M) antibody [EPR23526-89] 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)
This data was developed using Anti-Histone H3.3 (mutated K36M) antibody [EPR23526-89] ab254400, the same antibody clone in a different buffer formulation.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T cells labelling Histone H3 (mutated K36M) with Anti-Histone H3.3 (mutated K36M) antibody [EPR23526-89] 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.
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.
This data was developed using Anti-Histone H3.3 (mutated K36M) antibody [EPR23526-89] ab254400, the same antibody clone in a different buffer formulation.
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 Anti-Histone H3.3 (mutated K36M) antibody [EPR23526-89] 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.
This data was developed using Anti-Histone H3.3 (mutated K36M) antibody [EPR23526-89] ab254400, the same antibody clone in a different buffer formulation.
Dot blot analysis of Histone H3.3 (mutated K36M) using Anti-Histone H3.3 (mutated K36M) antibody [EPR23526-89] 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
This data was developed using Anti-Histone H3.3 (mutated K36M) antibody [EPR23526-89] ab254400, the same antibody clone in a different buffer formulation.
ELISA using Anti-Histone H3.3 (mutated K36M) antibody [EPR23526-89] 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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