Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Histone H3 (mutated K27M) antibody [EPR18340] - ChIP Grade – BSA and Azide free (ab240310)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR18340] to Histone H3 (mutated K27M) - ChIP Grade – BSA and Azide free
  • Suitable for: WB, ChIP, ICC/IF, IP, Indirect ELISA
  • Reacts with: Human

Overview

  • Product name

    Anti-Histone H3 (mutated K27M) antibody [EPR18340] - ChIP Grade – BSA and Azide free
    See all Histone H3 primary antibodies

  • Description

    Rabbit monoclonal [EPR18340] to Histone H3 (mutated K27M) - ChIP Grade – BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, ChIP, ICC/IF, IP, Indirect ELISAmore details
    Unsuitable for: IHC-P

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: His-tagged recombinant histone H3 K27M protein, His-tagged recombinant wild type histone H3 protein; HEK-293 transfected with Histone H3 (K27M) expression vector containing a myc-His-tag®, whole cell lysate. IP: HEK-293T transfected with myc-tagged H3 (K27M) expression vector whole cell lysate. ICC/IF: HEK-293T transfected with myc-tagged H3 (K27 mutated to M) expression vector. ChIP: Chromatin prepared from HEK-293T transfected with myc-tagged H3 (K27 mutated to M) expression vector.

  • General notes

    Ab240310 is the carrier-free version of ab190631. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with

    ab240310 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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 production
    For more information see here.

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

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

  • Form

    Liquid

  • Storage instructions

    Shipped at 4°C. Store at +4°C. Do Not Freeze.

  • Storage buffer

    pH: 7.2
    Constituent: PBS

  • Carrier free

    Yes
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EPR18340

  • Isotype

    IgG

Images

  • ChIP - Anti-Histone H3 (mutated K27M) antibody [EPR18340] - ChIP Grade – BSA and Azide free (ab240310)
    ChIP - Anti-Histone H3 (mutated K27M) antibody [EPR18340] - ChIP Grade – BSA and Azide free (ab240310)

    This data was developed using ab190631, the same antibody clone in a different buffer formulation.

    Chromatin was prepared from 293T transfected with myc-His tagged Histone H3(K27 M) and Histone H3 WT cells according to the Abcam Dual-X-ChIP protocol*. Cells were fixed with 1.5 mM EGS for 30mins and then formaldehyde for 10min. 
    The ChIP was performed with 25 µg of chromatin, 2 µg of ab190631 (red), 2 µg of ab213204 (red) (bottom panel, served as internal control) 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.

    *https://www.abcam.com/resources?keywords=X%20ChIP%20protocol

  • Immunoprecipitation - Anti-Histone H3 (mutated K27M) antibody [EPR18340] - ChIP Grade – BSA and Azide free (ab240310)
    Immunoprecipitation - Anti-Histone H3 (mutated K27M) antibody [EPR18340] - ChIP Grade – BSA and Azide free (ab240310)

    Histone H3 (K27 mutated to M) was immunoprecipitated from 0.35 mg 293T (human embryonic kidney epithelial cell) cells transfected with myc-tagged H3(K27M) expression vector whole cell lysate 10ug with ab190631 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab190631 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

    Lane 1: 293T (human embryonic kidney epithelial cell) cells transfected with myc-tagged H3(K27M) expression vector whole cell lysate 10ug

    Lane 2: ab190631 IP in 293T cells transfected with myc-tagged H3(K27M) expression vector whole cell lysate

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab190631 in 293T cells transfected with myc-tagged H3(K27M) expression vector whole cell lysate

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

    Exposure time: 5 seconds.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, azide, BSA and glycerol (ab190631).

  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (mutated K27M) antibody [EPR18340] - ChIP Grade – BSA and Azide free (ab240310)
    Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (mutated K27M) antibody [EPR18340] - ChIP Grade – BSA and Azide free (ab240310)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized 293T (human embryonic kidney epithelial cell) cells labelling Histone H3 (K27 mutated to M) with ab190631 at 1/5000 (0.2 ug/ml) dilution, followed by ab190631 anti- H3(K27 mutated to M) ab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1/1000 (2 ug/ml) dilution (Green). Confocal image showing nuclear staining in 293T cells transfected with myc-tagged H3 (K27 mutated to M) expression vector. is observed. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is Ab190631 anti- H3(K27 mutated to M) ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 (2 ug/ml) dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab190631).

  • Indirect ELISA - Anti-Histone H3 (mutated K27M) antibody [EPR18340] - ChIP Grade – BSA and Azide free (ab240310)
    Indirect ELISA - Anti-Histone H3 (mutated K27M) antibody [EPR18340] - ChIP Grade – BSA and Azide free (ab240310)

    This data was developed using ab190631, the same antibody clone in a different buffer formulation.

    ELISA using ab190631 at varying antibody concentrations and antigen concentration at 1000 ng/mL. An Alkaline Phosphatase-conjugated Goat Anti-Rabbit IgG (H+L) (1/2500) was used as the secondary antibody.

    The blue line indicates binding to the Histone H3 (mutated K27M) peptide. Binding to the following peptides was not seen:

    Histone H3 WT,
    Histone H3 (mono methyl K27),
    Histone H3 (di methyl K27),
    Histone H3 (tri methyl K27).

    This indicates the specificity of ab190631 for mutated K27M of Histone H3.

  • Anti-Histone H3 (mutated K27M) antibody [EPR18340] - ChIP Grade – BSA and Azide free (ab240310)
    Anti-Histone H3 (mutated K27M) antibody [EPR18340] - ChIP Grade – BSA and Azide free (ab240310)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Alternative products to Anti-Histone H3 (mutated K27M) antibody [EPR18340] - ChIP Grade – BSA and Azide free (ab240310)

© 2021 Astana Biomed Group LLP. All rights reserved.