Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Histone H3.3 (mutated G34R) antibody [EPR23519-91] - ChIP Grade – BSA and Azide free (ab273635)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR23519-91] to Histone H3.3 (mutated G34R) - ChIP Grade – BSA and Azide free
  • Suitable for: IHC-P, Dot blot, WB, Flow Cyt, ChIP, IP
  • Reacts with: Human

Overview

  • Product name

    Anti-Histone H3.3 (mutated G34R) antibody [EPR23519-91] - ChIP Grade – BSA and Azide free
    See all Histone H3.3 primary antibodies

  • Description

    Rabbit monoclonal [EPR23519-91] to Histone H3.3 (mutated G34R) - ChIP Grade – BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IHC-P, Dot blot, WB, Flow Cyt, ChIP, IPmore details
    Unsuitable for: ICC

  • Species reactivity

    Reacts with: Human

  • Immunogen

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

  • Positive control

    • WB: HEK-293T transfected with Histone H3.3 G34R expression vector containing a myc-His-tag®, whole cell lysate. IHC-P: Human giant tumor of bone tissue. Flow cyt: HEK-293T cells transfected with myc-tagged Histone H3.3 G34R expression vector. IP: HEK-293T transfected with Histone H3.3 G34R expression vector containing a myc-His-tag® whole cell lysate. ChIP: Chromatin prepared from HEK-293T transfected with Histone H3.3 (mutated G34 R).

  • General notes

    ab273635 is the carrier-free version of ab254402. 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.

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

    EPR23519-91

  • Isotype

    IgG

  • Research areas

Images

  • ChIP - Anti-Histone H3.3 (mutated G34R) antibody [EPR23519-91] - ChIP Grade – BSA and Azide free (ab273635)
    ChIP - Anti-Histone H3.3 (mutated G34R) antibody [EPR23519-91] - ChIP Grade – BSA and Azide free (ab273635)

    Chromatin was prepared from 293T transfacted with Histone H3.3(mutated G34 R) and 293T transfacted with Histone H3.3(WT)
    cells according to the Abcam X-ChIP protocol*. Cells were fixed with formaldehyde for 10min.
    The ChIP was performed with 25 µg of chromatin, 2 µg of ab254402 (red), or 2 µg of rabbit normal IgG ab172730 (gray) and 25 µl of Protein A/G Dynabeads. The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci).

    *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 (ab254402).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3.3 (mutated G34R) antibody [EPR23519-91] - ChIP Grade – BSA and Azide free (ab273635)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3.3 (mutated G34R) antibody [EPR23519-91] - ChIP Grade – BSA and Azide free (ab273635)

    Immunohistochemical analysis of paraffin-embedded Human giant cell tumor of bone tissue labeling Histone H3.3 (mutated G34 R) with ab254402 at 1/2000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on human giant cell tumor of bone (PMID: 29241742). The section was incubated with ab254402 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 a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

     

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab254402).

  • Western blot - Anti-Histone H3.3 (mutated G34R) antibody [EPR23519-91] - ChIP Grade – BSA and Azide free (ab273635)
    Western blot - Anti-Histone H3.3 (mutated G34R) antibody [EPR23519-91] - ChIP Grade – BSA and Azide free (ab273635)
    All lanes : Anti-Histone H3.3 (mutated G34R) antibody [EPR23519-91] - ChIP Grade (ab254402) at 1/1000 dilution

    Lane 1 : HEK-293T (human embryonic kidney) transfected with Histone H3.3 G34R expression vector containi a myc-His-tag®, whole cell lysate
    Lane 2 : HEK-293T transfected with Histone H3.3 (WT) expression vector containi a myc-His-tag®, whole cell lysate
    Lane 3 : HEK-293T (human embryonic kidney) transfected with Histone H3.3 G34V expression vector containi a myc-His-tag®, whole cell lysate
    Lane 4 : HEK-293T (human embryonic kidney) transfected with Histone H3.3 G34W expression vector containi a myc-His-tag®, whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution

    Predicted band size: 15 kDa
    Observed band size: 20 kDa
    why is the actual band size different from the predicted?



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

    Exposure time: 26 seconds . 

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab254402).

  • Flow Cytometry - Anti-Histone H3.3 (mutated G34R) antibody [EPR23519-91] - ChIP Grade – BSA and Azide free (ab273635)
    Flow Cytometry - Anti-Histone H3.3 (mutated G34R) antibody [EPR23519-91] - ChIP Grade – BSA and Azide free (ab273635)

    Flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized HEK-293T (Human embryonic kidney epithelial cell) (transfected with myc-tagged Histone H3.3 WT expression vector) (Left) or myc-tagged Histone H3.3 G34R expression vector (Right) cells labelling Histone H3.3 (mutated G34 R) with ab254402 at 1/5000 dilution (0.01ug) (Both panels). 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 (ab254402).

  • Immunoprecipitation - Anti-Histone H3.3 (mutated G34R) antibody [EPR23519-91] - ChIP Grade – BSA and Azide free (ab273635)
    Immunoprecipitation - Anti-Histone H3.3 (mutated G34R) antibody [EPR23519-91] - ChIP Grade – BSA and Azide free (ab273635)

    Histone H3.3 (mutated G34 R) was immunoprecipitated from 0.35 mg HEK-293T (human embryonic kidney) (transfected with Histone H3.3 G34R expression vector containing a myc-His-tag®) whole cell lysate with ab254402 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab254402 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

    Lane 1: HEK-293T (human embryonic kidney) transfected with Histone H3.3 G34R expression vector containing a myc-His-tag®, whole cell lysate 10 ug

    Lane 2: ab254402 IP in HEK-293T transfected with Histone H3.3 G34R expression vector containing a myc-His-tag® whole cell lysate

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab254402 in HEK-293T transfected with Histone H3.3 G34R expression vector containing a myc-His-tag® whole cell lysate

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

    Exposure time: 41 seconds.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab254402).

  • Dot Blot - Anti-Histone H3.3 (mutated G34R) antibody [EPR23519-91] - ChIP Grade – BSA and Azide free (ab273635)
    Dot Blot - Anti-Histone H3.3 (mutated G34R) antibody [EPR23519-91] - ChIP Grade – BSA and Azide free (ab273635)

    Dot blot analysis of Histone H3.3 (mutated G34 R) labeled with ab254402 at 1/1000 dilution.

    Lane 1: Histone H3.3 H3G34R peptide (aa28-40).
    Lane 2: Histone H3.3 H3G34R peptide (aa26-36).
    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 (ab254402).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3.3 (mutated G34R) antibody [EPR23519-91] - ChIP Grade – BSA and Azide free (ab273635)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3.3 (mutated G34R) antibody [EPR23519-91] - ChIP Grade – BSA and Azide free (ab273635)

    Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling Histone H3.3 (mutated G34 R) with ab254402 at 1/2000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Negative control: no staining on human testis. The section was incubated with ab254402 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 a 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 (ab254402).

  • Anti-Histone H3.3 (mutated G34R) antibody [EPR23519-91] - ChIP Grade – BSA and Azide free (ab273635)
    Anti-Histone H3.3 (mutated G34R) antibody [EPR23519-91] - ChIP Grade – BSA and Azide free (ab273635)

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