Anti-Histone H3 (tri methyl K27, phospho S28) antibody (ab108268)

Overview

  • Product nameAnti-Histone H3 (tri methyl K27, phospho S28) antibody
    See all Histone H3 primary antibodies
  • Description
    Rabbit polyclonal to Histone H3 (tri methyl K27, phospho S28)
  • SpecificityThis antibody detects Histone H3 that is both tri-methylated at K27 and phosphorylated at S28. It is believed that Histone H3 (tri methyl K27, phospho S28) is a double mark and that Histone H3 (tri-methyl K27) phosphorylation at S28 by the kinases MSK1 and MSK2 occurs in response to mitogenic, stress and differentiation signals leading to displacement of PcG proteins from target gene promoters.
  • Tested applicationsSuitable for: WBmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse, Rat, Chicken, Cow, Zebrafish
  • Immunogen

    Synthetic peptide corresponding to Human Histone H3 aa 1-100 (tri methyl K27, phospho S28) conjugated to Keyhole Limpet Haemocyanin (KLH).
    Database link: P68431

  • Positive control
    • This antibody gave a positive signal in HeLa Histone and Calf Thymus Histone Nuclear lysates.

Properties

  • FormLiquid
  • Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage bufferpH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: PBS

    Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
  • Concentration information loading...
  • PurityImmunogen affinity purified
  • ClonalityPolyclonal
  • IsotypeIgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab108268 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB Use a concentration of 1 µg/ml. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa).

Target

Anti-Histone H3 (tri methyl K27, phospho S28) antibody images

  • All lanes : Anti-Histone H3 (tri methyl K27, phospho S28) antibody (ab108268) at 1 µg/ml

    Lane 1 : HeLa Histone Preparation Nuclear Lysate at 10 µg
    Lane 2 : Calf Thymus Histone Preparation Nuclear Lysate at 0.5 µg
    Lane 3 : HeLa Histone Preparation Nuclear Lysate at 10 µg with Human Histone H3 (tri methyl K27, phospho S28) peptide (ab173750) at 1 µg/ml
    Lane 4 : Calf Thymus Histone Preparation Nuclear Lysate at 0.5 µg with Human Histone H3 (tri methyl K27, phospho S28) peptide (ab173750) at 1 µg/ml
    Lane 5 : HeLa Histone Preparation Nuclear Lysate at 10 µg with Human Histone H3 peptide (ab173779) at 1 µg/ml
    Lane 6 : Calf Thymus Histone Preparation Nuclear Lysate at 0.5 µg with Human Histone H3 peptide (ab173779) at 1 µg/ml
    Lane 7 : HeLa Histone Preparation Nuclear Lysate at 10 µg with Human Histone H3 (tri methyl K27) peptide (ab1782) at 1 µg/ml
    Lane 8 : Calf Thymus Histone Preparation Nuclear Lysate at 0.5 µg with Human Histone H3 (tri methyl K27) peptide (ab1782) at 1 µg/ml
    Lane 9 : HeLa Histone Preparation Nuclear Lysate at 10 µg with Human Histone H3 (phospho S28) peptide (ab14793) at 1 µg/ml
    Lane 10 : Calf Thymus Histone Preparation Nuclear Lysate at 0.5 µg with Human Histone H3 (phospho S28) peptide (ab14793) at 1 µg/ml

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 15 kDa
    Observed band size : 17 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 100 kDa (possible non-specific binding),52 kDa (possible non-specific binding).

    Exposure time : 8 minutes

    This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Bovine Serum Albumin before being incubated with ab108268 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.

  • ICC/IF Peptide competition assay performed on Methanol fixed HeLa cells. The cells were 100% Methanol fixed for 5 minutes at Room temperature and then incubated in 1%BSA / 10% normal Goat serum / 0.3M glycine in 0.1% PBS-TWEEN for 1hour at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The following peptides were chosen and incubated with ab108268 for 1 hour at room temperature, (non-mod block ab187169, Single modified block (tri-methyl-k27) ab1782, Single modified block (phosphor-S28) ab14793, Double modified block (K27 + S28) ab173822. ab108268 was used at 1ug/ml. Peptides were incubated at twice the concentration of the primary antibody. The cells were then incubated with either the peptide-competed antibody or the control antibody without peptide overnight at +4°C. The secondary antibody (pseudo-colored green) was Goat anti-Rabbit (ab150081) IgG (H+L) preadsorbed used at a 1/1000 dilution for 1hour at room temperature. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.

References for Anti-Histone H3 (tri methyl K27, phospho S28) antibody (ab108268)

ab108268 has not yet been referenced specifically in any publications.

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