Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Histone H3.3 (phospho S31) antibody [EPR1873] (ab92628)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR1873] to Histone H3.3 (phospho S31)
  • Suitable for: Flow Cyt, Dot blot, WB, IHC-P, ICC/IF
  • Reacts with: Human

Overview

  • Product name

    Anti-Histone H3.3 (phospho S31) antibody [EPR1873]
    See all Histone H3.3 primary antibodies

  • Description

    Rabbit monoclonal [EPR1873] to Histone H3.3 (phospho S31)

  • Host species

    Rabbit

  • Specificity

    Detects Histone H3.3 when phosphorylated at S31.

  • Tested Applications & Species

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

  • Immunogen

    Synthetic peptide within Human Histone H3.3 aa 1-100 (phospho S31). The exact sequence is proprietary.
    Database link: P84243

  • Positive control

    • Untreated Hela cells and HeLa cell lysate treated with Calyculin A and/or nocodazole. Human cervical carcinoma

  • General notes

     

     

    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.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

  • Form

    Liquid

  • Storage instructions

    Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.

  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.5% BSA
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EPR1873

  • Isotype

    IgG

  • Research areas

Images

  • Western blot - Anti-Histone H3.3 (phospho S31) antibody [EPR1873] (ab92628)
    Western blot - Anti-Histone H3.3 (phospho S31) antibody [EPR1873] (ab92628)
    All lanes : Anti-Histone H3.3 (phospho S31) antibody [EPR1873] (ab92628) at 1/1000 dilution

    Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 2 : Whole cell lysate from HeLa (Human epithelial cell line from cervix adenocarcinoma) cells treated with 100ng/ml of nocodazole for 18 hours
    Lane 3 : Whole cell lysate from HeLa (Human epithelial cell line from cervix adenocarcinoma) cells treated with 100ng/ml of nocodazole for 18 hours. Membrane incubated with phosphatase

    Lysates/proteins at 15 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 15 kDa


    Exposure time: 15 seconds


    Blocking/dilution buffer: 2% BSA/TBST

  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3.3 (phospho S31) antibody [EPR1873] (ab92628)
    Immunocytochemistry/ Immunofluorescence - Anti-Histone H3.3 (phospho S31) antibody [EPR1873] (ab92628)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% tritonX-100 HeLa (Human epithelial cell line from cervix adenocarcinoma) cells, LP starved and non-starved, labeling anti-Histone H3.3 (phospho S31) with Ab92628 at 1/500 dilution followed by Goat anti-Rabbit secondary IgG AlexaFluor®488 (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing nuclear staining on M phase of HeLa cells, then the signal decreased after LP treatment.

    For the pan antibody, there was no great difference after LP treatment. The data showed mostly nuclear staining.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3.3 (phospho S31) antibody [EPR1873] (ab92628)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3.3 (phospho S31) antibody [EPR1873] (ab92628)

    Paraffin-embedded human cervical carcinoma labelled with ab92628 at 1/100 dilution.

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Flow Cytometry - Anti-Histone H3.3 (phospho S31) antibody [EPR1873] (ab92628)
    Flow Cytometry - Anti-Histone H3.3 (phospho S31) antibody [EPR1873] (ab92628)

    Flow Cytometry analysis of Hela (human cervix adenocarcinoma) cells treated with 100 ng/ml Nocodazole for 18 hours, labeling Histone H3.3 with purified ab92628 at 1/150 dilution (10µg/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti-rabbit IgG (Alexa Fluor® 488) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control. Untreated control - Hela (human cervix adenocarcinoma) cells untreated with 100 ng/ml Nocodazole for 18 hours (Green).

  • Dot Blot - Anti-Histone H3.3 (phospho S31) antibody [EPR1873] (ab92628)
    Dot Blot - Anti-Histone H3.3 (phospho S31) antibody [EPR1873] (ab92628)

    Dot Blot analysis of Lane 1: Histone H3.3 (pS31) phospho peptide and Lane 2: Histone H3.3 non-phospho peptide labeling Histone H3.3 (phospho S31) with ab92628 at 1/1000 dilution. 5% NFDM/TBST was used as the diluting and blocking buffer. 

    ab97051 Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated was used as the secondary antibody at 1/100000 dilution. Exposure time: 10 seconds.

     

     

     

     

     

  • Western blot - Anti-Histone H3.3 (phospho S31) antibody [EPR1873] (ab92628)
    Western blot - Anti-Histone H3.3 (phospho S31) antibody [EPR1873] (ab92628)
    All lanes : Anti-Histone H3.3 (phospho S31) antibody [EPR1873] (ab92628) at 1/1000 dilution

    Lane 1 : HeLa cell lysate untreated
    Lane 2 : HeLa cell lysate treated with Calyculin A

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

    Predicted band size: 15 kDa

  • Western blot - Anti-Histone H3.3 (phospho S31) antibody [EPR1873] (ab92628)
    Western blot - Anti-Histone H3.3 (phospho S31) antibody [EPR1873] (ab92628) This image is courtesy of an anonymous Abreview
    Representative western blot detecting Histone H3.3 (phospho S31) using ab92628 at 1/3000 dilution.

    Recombinant Histone H3.3 protein was treated with recombinant kinase and 200 µM ATP for the indicated times. 0.2 µg of protein was loaded to each lane and phosphorylated Histone H3.3 was detected using ab92628. An HRP-conjugated goat anti-rabbit polyclonal (1/20000) was used as the secondary antibody

    See Abreview

  • Anti-Histone H3.3 (phospho S31) antibody [EPR1873] (ab92628)
    Anti-Histone H3.3 (phospho S31) antibody [EPR1873] (ab92628)

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