Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Histone H3 (citrulline R8) antibody [EPR20358-13] - BSA and Azide free (ab232939)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR20358-13] to Histone H3 (citrulline R8) - BSA and Azide free
  • Suitable for: ICC, ELISA, WB, Flow Cyt, IP
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-Histone H3 (citrulline R8) antibody [EPR20358-13] - BSA and Azide free
    See all Histone H3 primary antibodies

  • Description

    Rabbit monoclonal [EPR20358-13] to Histone H3 (citrulline R8) - BSA and Azide free

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    IP
    Human
    WB
    Rat
    See all applications and species data

  • Immunogen

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

  • Positive control

    • ICC/IF: HEK-293T cells transfected with GFP-tagged PADI4, then treated with 10 mM CaCl2. IP: HEK-293T transfected with PADI4 (WT), then treated with 10mM CaCl2 and 10µM Ionomycin. Flow Cyt: HEK-293T transfected with PADI4 (WT), then treated with 10mM CaCl2 and 10µM Ionomycin.

  • General notes

    Ab232939 is the carrier-free version of ab219406. 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

    ab232939 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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

    EPR20358-13

  • Isotype

    IgG

  • Research areas

Images

  • Immunoprecipitation - Anti-Histone H3 (citrulline R8) antibody [EPR20358-13] - BSA and Azide free (ab232939)
    Immunoprecipitation - Anti-Histone H3 (citrulline R8) antibody [EPR20358-13] - BSA and Azide free (ab232939)

    Histone H3 (citrulline R8) was immunoprecipitated from 0.35 mg of HEK-293T (Human epithelial cell line from embryonic kidney) transfected with PADI4 (WT), then treated with 10mM CaCl2 and 10μM Ionomycin for 2 hours, whole cell lysate with ab219406 at 1/30 dilution.

    Western blot was performed from the immunoprecipitate using ab219406 at 1/1000 dilution.

    VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: HEK-293T transfected with PADI4 (WT), then treated with 10mM CaCl2 and 10μM Ionomycin for 2 hours, whole cell lysate 10 µg (Input).

    Lane 2: ab219406 IP in HEK-293T transfected with PADI4 (WT), then treated with 10mM CaCl2 and 10μM Ionomycin for 2 hours, whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab219406 in HEK-293T transfected with PADI4 (WT), then treated with 10mM CaCl2 and 10μM Ionomycin for 2 hours, whole cell lysate.

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

    Exposure time: 1 second.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab219406).

  • ELISA - Anti-Histone H3 (citrulline R8) antibody [EPR20358-13] - BSA and Azide free (ab232939)
    ELISA - Anti-Histone H3 (citrulline R8) antibody [EPR20358-13] - BSA and Azide free (ab232939)

    Direct ELISA using ab219406 at 0-1000 ng/ml, followed by Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) at 1/2500 dilution. Antigen concentration: 1000 ng/ml.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab219406).

  • Flow Cytometry - Anti-Histone H3 (citrulline R8) antibody [EPR20358-13] - BSA and Azide free (ab232939)
    Flow Cytometry - Anti-Histone H3 (citrulline R8) antibody [EPR20358-13] - BSA and Azide free (ab232939)

    Flow cytometric analysis of 4% paraformaldehyde-fixed HEK-293T (Human epithelial cell line from embryonic kidney) transfected with empty vector (left panel) or PADI4 (WT, right panel), then treated with 10mM CaCl2 and 10μM Ionomycin for 2 hours, labeling Histone H3 (citrulline R8) with ab219406 at 1/500 dilution, followed by Goat anti rabbit IgG (Alexa Fluor® 647) at 1/2000 dilution.

    Positive signal is obtained from HEK-293T cells transfected with WT PADI4 treated with 10mM CaCl2 and 10μM Ionomycin for 2 hours.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab219406).

  • Western blot - Anti-Histone H3 (citrulline R8) antibody [EPR20358-13] - BSA and Azide free (ab232939)
    Western blot - Anti-Histone H3 (citrulline R8) antibody [EPR20358-13] - BSA and Azide free (ab232939)
    All lanes : Anti-Histone H3 (citrulline R8) antibody [EPR20358-13] - BSA and Azide free (ab232939) at 1/1000 dilution

    Lane 1 : E12 mouse embryo lysates
    Lane 2 : E12 rat embryo lysates

    Lysates/proteins at 15 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

    Predicted band size: 15 kDa


    Exposure time: 6 seconds
  • Immunocytochemistry - Anti-Histone H3 (citrulline R8) antibody [EPR20358-13] - BSA and Azide free (ab232939)
    Immunocytochemistry - Anti-Histone H3 (citrulline R8) antibody [EPR20358-13] - BSA and Azide free (ab232939)

    Clone EPR20358-13 (ab232939) has been successfully conjugated by Abcam. This image was generated using Anti-Histone H3 (citrulline R8) antibody [EPR20358-13] (Alexa Fluor® 647). Please refer to ab237370 for protocol details.

    Immunofluorescent analysis of 4% PFA-fixed, 0.1% Triton X-100 permeabilized HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) cells labeling Histone H3 with ab237370 at 1/100 dilution (red). Confocal image showing nuclear staining in HEK-293T cells transfected with a GFP-tagged PADI4 expression construct (green), then treated with 10mM CaCl2 for 2 hours and 10µM Ionomycin for 2 hours. Nuclear counterstain is DAPI (blue).

  • Immunocytochemistry - Anti-Histone H3 (citrulline R8) antibody [EPR20358-13] - BSA and Azide free (ab232939)
    Immunocytochemistry - Anti-Histone H3 (citrulline R8) antibody [EPR20358-13] - BSA and Azide free (ab232939)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HEK-293T (Human epithelial cell line from embryonic kidney) cells transfected with GFP only or a GFP-tagged PADI4 expression construct, then treated with 10mM CaCl2 for 2 hours, labeling Histone H3 (citrulline R8) with ab219406 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing positive staining on HEK-293T cells transfected with a GFP-tagged PADI4 expression construct, then treated with 10 mM CaCl2 for 2 hours. The nuclear counterstain is DAPI (blue).

    Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab219406).

  • Anti-Histone H3 (citrulline R8) antibody [EPR20358-13] - BSA and Azide free (ab232939)
    Anti-Histone H3 (citrulline R8) antibody [EPR20358-13] - BSA and Azide free (ab232939)

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