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AB270796

Anti-Histone H3 (mutated K36M) antibody [EPR23614-91] - BSA and Azide free

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Rabbit Recombinant Monoclonal H3 mutated K36M antibody. Carrier free. Suitable for ChIC/CUT&RUN-seq, IP, WB, IHC-P, I-ELISA, ICC/IF, Flow Cyt (Intra), IHC-Fr and reacts with Transfected cell line - Human, Human, Transfected cell lysate - Human, Synthetic peptide samples.

View Alternative Names

H3FA, HIST1H3A, H3C2, H3FL, HIST1H3B, H3C3, H3FC HIST1H3C, H3C4, H3FB, HIST1H3D, H3C6, H3FD, HIST1H3E, H3C7, H3FI, HIST1H3F, H3C8, H3FH, HIST1H3G, H3C10, H3FK, HIST1H3H, H3C11, H3FF, HIST1H3I, H3C12, H3FJ, HIST1H3J, HIST1H3C, H3FC, H3C1, Histone H3.1, Histone H3/a, Histone H3/b, Histone H3/c, Histone H3/d, Histone H3/f, Histone H3/h, Histone H3/i, Histone H3/j, Histone H3/k, Histone H3/l, H3K36M, H3K36, H3.3A, H3F3, H3F3A, PP781, H3-3B, H3.3B, H3F3B, H3-3A, Histone H3.3, H3K36M, H3.3K36M, H3K36

7 Images
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (mutated K36M) antibody [EPR23614-91] - BSA and Azide free (AB270796)
  • IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (mutated K36M) antibody [EPR23614-91] - BSA and Azide free (AB270796)

Immunohistochemical analysis of paraffin-embedded Human chondroblastoma tissue labeling Histone H3 (mutated K36 M) with ab256384 at 1/4000 dilution followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Nuclear staining in human chondroblastoma (PMID : 29757500). Counterstained with Hematoxylin.

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

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab256384).

Flow Cytometry (Intracellular) - Anti-Histone H3 (mutated K36M) antibody [EPR23614-91] - BSA and Azide free (AB270796)
  • Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Histone H3 (mutated K36M) antibody [EPR23614-91] - BSA and Azide free (AB270796)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized 293T transfected with myc tagged Histone H3 construct (Left) or myc tagged Histone H3 K36M construct (Right) cells labelling Histone H3 (mutated K36 M) with ab256384 at 1/500 compared with a isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 was used as the secondary antibody.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab256384).

Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (mutated K36M) antibody [EPR23614-91] - BSA and Azide free (AB270796)
  • ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (mutated K36M) antibody [EPR23614-91] - BSA and Azide free (AB270796)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HEK-293 cells labelling Histone H3 (mutated K36 M) with ab256384 at 1/50 dilution, followed by a secondary antibody at 1/1000 dilution (Green). Confocal image showing nuclear staining in HEK-293 cell line transfected with myc-tagged Histone H3 K36M expression vector. 2233S Myc-Tag (9B11) Mouse mAb (Alexa Fluor® 647 Conjugate) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab256384).

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (mutated K36M) antibody [EPR23614-91] - BSA and Azide free (AB270796)
  • IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (mutated K36M) antibody [EPR23614-91] - BSA and Azide free (AB270796)

Immunohistochemical analysis of paraffin-embedded Human giant cell tumor of bone tissue labeling Histone H3 (mutated K36 M) with ab256384 at 1/4000 dilution followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Negative control : No staining in human giant cell tumor of bone (PMID : 29757500). Counterstained with Hematoxylin.

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

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab256384).

Immunoprecipitation - Anti-Histone H3 (mutated K36M) antibody [EPR23614-91] - BSA and Azide free (AB270796)
  • IP

Unknown

Immunoprecipitation - Anti-Histone H3 (mutated K36M) antibody [EPR23614-91] - BSA and Azide free (AB270796)

Histone H3.3 (mutated K36 M) was immunoprecipitated from 0.35 mg HEK-293 transfected with Histone H3.3 K36M (mutate) expression vector containing a myc-His-tag®, whole cell lysate with ab256384 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab256384 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : HEK-293 transfected with Histone H3.3 K36M (mutate) expression vector containing a myc-His-tag®, whole cell lysate 10 ug

Lane 2 : ab256384 IP in HEK-293 transfected with Histone H3.3 K36M (mutate) expression vector containing a myc-His-tag®, whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab256384 in HEK-293 transfected with Histone H3.3 K36M (mutate) expression vector containing a myc-His-tag®, whole cell lysate

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

Exposure time : 10 seconds

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab256384).

All lanes:

Immunoprecipitation - Anti-Histone H3 (mutated K36M) antibody [EPR23614-91] (<a href='/en-us/products/primary-antibodies/histone-h3-mutated-k36m-antibody-epr23614-91-ab256384'>ab256384</a>)

Predicted band size: 15 kDa

false

ChIC/CUT&RUN sequencing - Anti-Histone H3 (mutated K36M) antibody [EPR23614-91] - BSA and Azide free (AB270796)
  • ChIC/CUT&RUN-seq

Supplier Data

ChIC/CUT&RUN sequencing - Anti-Histone H3 (mutated K36M) antibody [EPR23614-91] - BSA and Azide free (AB270796)

ChIC/CUT&RUN was performed using a pAG-MNAse at a final concentration of 700 ng/mL, 2.5 x 10^5 293T cells transfected with a H3.3K36M (Mutant) or H3.3 (WT) plasmid and 3µg of ab256384 [EPR23614-91]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown. Additional screenshots of mapped reads can be found in the Protocol booklet in the Product Protocol section. The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods. This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab256384).

Indirect ELISA - Anti-Histone H3 (mutated K36M) antibody [EPR23614-91] - BSA and Azide free (AB270796)
  • I-ELISA

Lab

Indirect ELISA - Anti-Histone H3 (mutated K36M) antibody [EPR23614-91] - BSA and Azide free (AB270796)

This data was developed using ab256384, the same antibody clone in a different buffer formulation.

Indirect ELISA using ab256384 at varying antibody concentrations and antigen concentration at 100 ng/mL. An Alkaline Phosphatase-conjugated Goat Anti-Rabbit IgG (H+L) (1/2500) was used as the secondary antibody.

Binding was seen for Histone H3 (mutated K36M) peptide. Binding to the following peptides was not seen :

Histone H3 WT,
Histone H3 (mono methyl K36),
Histone H3 (di methyl K36),
Histone H3 (tri methyl K36).

This indicates the specificity of ab256384 for mutated K36M of Histone H3.

  • Unconjugated

    Anti-Histone H3 (mutated K36M) antibody [EPR23614-91]

  • 660 APC

    APC Anti-Histone H3 (mutated K36M) antibody [EPR23614-91]

  • 519 Alexa Fluor® 488

    Alexa Fluor® 488 Anti-Histone H3 (mutated K36M) antibody [EPR23614-91]

  • 565 Alexa Fluor® 555

    Alexa Fluor® 555 Anti-Histone H3 (mutated K36M) antibody [EPR23614-91]

  • 603 Alexa Fluor® 568

    Alexa Fluor® 568 Anti-Histone H3 (mutated K36M) antibody [EPR23614-91]

  • 617 Alexa Fluor® 594

    Alexa Fluor® 594 Anti-Histone H3 (mutated K36M) antibody [EPR23614-91]

  • 665 Alexa Fluor® 647

    Alexa Fluor® 647 Anti-Histone H3 (mutated K36M) antibody [EPR23614-91]

  • 775 Alexa Fluor® 750

    Alexa Fluor® 750 Anti-Histone H3 (mutated K36M) antibody [EPR23614-91]

  • HRP

    HRP Anti-Histone H3 (mutated K36M) antibody [EPR23614-91]

  • 578 PE

    PE Anti-Histone H3 (mutated K36M) antibody [EPR23614-91]

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR23614-91

Isotype

IgG

Carrier free

Yes

Reacts with

Human

Applications

IHC-Fr, Flow Cyt (Intra), ICC/IF, WB, I-ELISA, ChIC/CUT&RUN-seq, IHC-P, IP

applications

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "ChICCUTRUNseq" : {"fullname" : "ChIC/CUT&RUN sequencing", "shortname":"ChIC/CUT&RUN-seq"}, "IP" : {"fullname" : "Immunoprecipitation", "shortname":"IP"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "IHCP" : {"fullname" : "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)", "shortname":"IHC-P"}, "IELISA" : {"fullname" : "Indirect ELISA", "shortname":"I-ELISA"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"}, "FlowCytIntra" : {"fullname" : "Flow Cytometry (Intracellular)", "shortname":"Flow Cyt (Intra)"}, "ChIP" : {"fullname" : "ChIP", "shortname":"ChIP"}, "IHCFr" : {"fullname" : "Immunohistochemistry (Frozen sections)", "shortname":"IHC-Fr"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "ChICCUTRUNseq-species-checked": "guaranteed", "ChICCUTRUNseq-species-dilution-info": "", "ChICCUTRUNseq-species-notes": "", "IP-species-checked": "guaranteed", "IP-species-dilution-info": "", "IP-species-notes": "", "WB-species-checked": "guaranteed", "WB-species-dilution-info": "", "WB-species-notes": "", "IHCP-species-checked": "testedAndGuaranteed", "IHCP-species-dilution-info": "", "IHCP-species-notes": "<p>Heat mediated antigen retrieval using <a href='/en-us/products/buffers/antigen-retrieval-buffer-100x-tris-edta-buffer-ph-90-ab93684'>ab93684</a> (Tris/EDTA buffer, pH 9.0)</p>", "IELISA-species-checked": "guaranteed", "IELISA-species-dilution-info": "", "IELISA-species-notes": "", "ICCIF-species-checked": "testedAndGuaranteed", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "<p></p>", "FlowCytIntra-species-checked": "guaranteed", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "", "ChIP-species-checked": "notRecommended", "ChIP-species-dilution-info": "", "ChIP-species-notes": "<p></p>", "IHCFr-species-checked": "guaranteed", "IHCFr-species-dilution-info": "", "IHCFr-species-notes": "<p></p>" }, "Mouse": { "ChICCUTRUNseq-species-checked": "notRecommended", "ChICCUTRUNseq-species-dilution-info": "", "ChICCUTRUNseq-species-notes": "", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "", "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "<p>Heat mediated antigen retrieval using <a href='/en-us/products/buffers/antigen-retrieval-buffer-100x-tris-edta-buffer-ph-90-ab93684'>ab93684</a> (Tris/EDTA buffer, pH 9.0)</p>", "IELISA-species-checked": "notRecommended", "IELISA-species-dilution-info": "", "IELISA-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "FlowCytIntra-species-checked": "notRecommended", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "", "ChIP-species-checked": "notRecommended", "ChIP-species-dilution-info": "", "ChIP-species-notes": "", "IHCFr-species-checked": "notRecommended", "IHCFr-species-dilution-info": "", "IHCFr-species-notes": "" }, "Rat": { "ChICCUTRUNseq-species-checked": "notRecommended", "ChICCUTRUNseq-species-dilution-info": "", "ChICCUTRUNseq-species-notes": "", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "", "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "<p>Heat mediated antigen retrieval using <a href='/en-us/products/buffers/antigen-retrieval-buffer-100x-tris-edta-buffer-ph-90-ab93684'>ab93684</a> (Tris/EDTA buffer, pH 9.0)</p>", "IELISA-species-checked": "notRecommended", "IELISA-species-dilution-info": "", "IELISA-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "FlowCytIntra-species-checked": "notRecommended", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "", "ChIP-species-checked": "notRecommended", "ChIP-species-dilution-info": "", "ChIP-species-notes": "", "IHCFr-species-checked": "notRecommended", "IHCFr-species-dilution-info": "", "IHCFr-species-notes": "" }, "Synthetic peptide": { "ChICCUTRUNseq-species-checked": "notRecommended", "ChICCUTRUNseq-species-dilution-info": "", "ChICCUTRUNseq-species-notes": "", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "", "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "IELISA-species-checked": "testedAndGuaranteed", "IELISA-species-dilution-info": "", "IELISA-species-notes": "<p></p>", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "FlowCytIntra-species-checked": "notRecommended", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "", "ChIP-species-checked": "notRecommended", "ChIP-species-dilution-info": "", "ChIP-species-notes": "", "IHCFr-species-checked": "notRecommended", "IHCFr-species-dilution-info": "", "IHCFr-species-notes": "" }, "Transfected cell line - Human": { "ChICCUTRUNseq-species-checked": "testedAndGuaranteed", "ChICCUTRUNseq-species-dilution-info": "", "ChICCUTRUNseq-species-notes": "<p></p>", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "", "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "IELISA-species-checked": "notRecommended", "IELISA-species-dilution-info": "", "IELISA-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "FlowCytIntra-species-checked": "notRecommended", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "", "ChIP-species-checked": "notRecommended", "ChIP-species-dilution-info": "", "ChIP-species-notes": "", "IHCFr-species-checked": "notRecommended", "IHCFr-species-dilution-info": "", "IHCFr-species-notes": "" }, "Transfected cell lysate - Human": { "ChICCUTRUNseq-species-checked": "notRecommended", "ChICCUTRUNseq-species-dilution-info": "", "ChICCUTRUNseq-species-notes": "", "IP-species-checked": "testedAndGuaranteed", "IP-species-dilution-info": "", "IP-species-notes": "<p></p>", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "", "WB-species-notes": "<p></p>", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "IELISA-species-checked": "notRecommended", "IELISA-species-dilution-info": "", "IELISA-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "FlowCytIntra-species-checked": "testedAndGuaranteed", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "<p></p>", "ChIP-species-checked": "notRecommended", "ChIP-species-dilution-info": "", "ChIP-species-notes": "", "IHCFr-species-checked": "notRecommended", "IHCFr-species-dilution-info": "", "IHCFr-species-notes": "" } } }
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Product details

ab270796 is the carrier-free version of ab256384.

Patented technology
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

What are the advantages of a recombinant monoclonal antibody?
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.

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. 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.

Compatibility
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.

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Properties and storage information

Form
Liquid
Purification technique
Affinity purification Protein A
Storage buffer
pH: 7.2 - 7.4 Constituents: PBS
Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
Do Not Freeze
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Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:
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Target data

Core component of nucleosome. 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.
See full target information H3C1 mutated K36M

Additional targets

H3-3A mutated K36M
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chicCutRunSequencingBooklet
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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

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