Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Histone H3.3 (mutated G34V) antibody [EPR23520-5] - ChIP Grade – BSA and Azide free (ab272942)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR23520-5] to Histone H3.3 (mutated G34V) - ChIP Grade – BSA and Azide free
  • Suitable for: ICC/IF, Flow Cyt, IHC-P, Dot blot, IP, WB, ChIP
  • Reacts with: Human

Overview

  • Product name

    Anti-Histone H3.3 (mutated G34V) antibody [EPR23520-5] - ChIP Grade – BSA and Azide free
    See all Histone H3.3 primary antibodies

  • Description

    Rabbit monoclonal [EPR23520-5] to Histone H3.3 (mutated G34V) - ChIP Grade – BSA and Azide free

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    ChIP
    Human
    Flow Cyt
    Human
    ICC/IF
    Human
    IHC-P
    Human
    IP
    Human
    See all applications and species data

  • Immunogen

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

  • Positive control

    • WB: HEK-293T transfected with Histone H3.3 G34V expression vector containing a myc-His-tag, whole cell lysate. IHC-P: Human giant tumor of bone. ICC/IF: 293T cells transfected with Histone H3.3 H3G34V-Myc plasmid. Flow Cyt: 293T transfected myc-tagged Histone H3.3 H3G34V construct. IP: HEK-293T transfected with Histone H3.3 G34V expression vector containing a myc-His-tag whole cell lysate.

  • General notes

    ab272942 is the carrier-free version of ab254401. 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

    This product 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.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

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

    EPR23520-5

  • Isotype

    IgG

  • Research areas

Images

  • ChIP - Anti-Histone H3.3 (mutated G34V) antibody [EPR23520-5] - ChIP Grade – BSA and Azide free (ab272942)
    ChIP - Anti-Histone H3.3 (mutated G34V) antibody [EPR23520-5] - ChIP Grade – BSA and Azide free (ab272942)

    Chromatin was prepared from 293T transfected with myc-His tagged Histone H3.3 mutated G34V and Histone H3.3 WT cells according to the Abcam Dual-X-ChIP protocol*. Cells were fixed with then formaldehyde for 10min.
    The ChIP was performed with 25 µg of chromatin, 2 µg of ab254401 (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.

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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab254401).

  • Immunoprecipitation - Anti-Histone H3.3 (mutated G34V) antibody [EPR23520-5] - ChIP Grade – BSA and Azide free (ab272942)
    Immunoprecipitation - Anti-Histone H3.3 (mutated G34V) antibody [EPR23520-5] - ChIP Grade – BSA and Azide free (ab272942)

    Histone H3.3(mutated G34 V) was immunoprecipitated from 0.35 mg HEK-293T (human embryonic kidney) transfected with Histone H3.3 G34V expression vector containing a myc-His-tag® whole cell lysate 10 µg with ab254401 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab254401 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

    Lane 1: HEK-293T transfected with Histone H3.3 G34V expression vector containing a myc-His-tag® whole cell lysate 10 µg

    Lane 2: ab254401 IP in HEK-293T transfected with Histone H3.3 G34V expression vector containing a myc-His-tag® whole cell lysate

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab254401 in HEK-293T transfected with Histone H3.3 G34V expression vector containing a myc-His-tag® whole cell lysate

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

    Exposure time: 32 seconds.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab254401).

  • Flow Cytometry - Anti-Histone H3.3 (mutated G34V) antibody [EPR23520-5] - ChIP Grade – BSA and Azide free (ab272942)
    Flow Cytometry - Anti-Histone H3.3 (mutated G34V) antibody [EPR23520-5] - ChIP Grade – BSA and Azide free (ab272942)

    Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized 293T (Human embryonic kidney epithelial cell) transfected with myc tagged Histone H3.3 WT construct (Left panel) and myc-tagged Histone H3.3 H3G34V construct (Right panel) cells labelling Histone H3.3(mutated G34 V) with ab254401 at 1/500 dilution (0.1µg). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution 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 (ab254401).

  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3.3 (mutated G34V) antibody [EPR23520-5] - ChIP Grade – BSA and Azide free (ab272942)
    Immunocytochemistry/ Immunofluorescence - Anti-Histone H3.3 (mutated G34V) antibody [EPR23520-5] - ChIP Grade – BSA and Azide free (ab272942)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized 293T cells labelling Histone H3.3(mutated G34 V) with ab254401 at 1/1000 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 293T cells transfected with Histone H3.3 H3G34V-Myc plasmid, while no staining in 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab254401).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3.3 (mutated G34V) antibody [EPR23520-5] - ChIP Grade – BSA and Azide free (ab272942)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3.3 (mutated G34V) antibody [EPR23520-5] - ChIP Grade – BSA and Azide free (ab272942)

    Immunohistochemical analysis of paraffin-embedded human giant cell tumor of bone tissue labeling Histone H3.3(mutated G34 V) with ab254401 at 1/1000 dilution (0.542 µg/ml) dilution followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human giant cell tumor of bone. (PMID: 29241742). The section was incubated with ab254401 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab254401).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3.3 (mutated G34V) antibody [EPR23520-5] - ChIP Grade – BSA and Azide free (ab272942)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3.3 (mutated G34V) antibody [EPR23520-5] - ChIP Grade – BSA and Azide free (ab272942)

    Immunohistochemical analysis of paraffin-embedded human chondroblastoma tissue labeling Histone H3.3(mutated G34 V) with ab254401 at 1/1000 dilution (0.542 µg/ml) dilution followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Negative control: No staining on human chondroblastoma (PMID: 29241742).

    The section was incubated with ab254401 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab254401).

  • Dot Blot - Anti-Histone H3.3 (mutated G34V) antibody [EPR23520-5] - ChIP Grade – BSA and Azide free (ab272942)
    Dot Blot - Anti-Histone H3.3 (mutated G34V) antibody [EPR23520-5] - ChIP Grade – BSA and Azide free (ab272942)

    Dot blot analysis of Histone H3.3 (mutated G34 V) labeled with ab254401 at 1/1000 dilution.

    Lane 1: Histone H3.3 H3G34V peptide (aa28-40).
    Lane 2: Histone H3.3 H3G34V peptide (aa26-38).
    Lane 3: Histone H3.3 WT peptide (aa26-40).

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab254401).

  • Anti-Histone H3.3 (mutated G34V) antibody [EPR23520-5] - ChIP Grade – BSA and Azide free (ab272942)
    Anti-Histone H3.3 (mutated G34V) antibody [EPR23520-5] - ChIP Grade – BSA and Azide free (ab272942)

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