Anti-phosphohistidine antibody (ab231709)

Overview

  • Product name

    Anti-phosphohistidine antibody
  • Description

    Sheep polyclonal to phosphohistidine
  • Host species

    Sheep
  • Tested applications

    Suitable for: WB, IP, Competitive ELISAmore details
  • Species reactivity

    Reacts with: Species independent
  • Immunogen

    Other Immunogen Type. A non-hydrolysable analogue of phosphohistidine, 1-(2-aminoethyl)pyrazol-4-ylphosphonic acid, conjugated to keyhole limpet hemocyanin (KLH) via a glutaraldehyde linkage.

  • Positive control

    • Competitive ELISA: Phosphorylated amino acids and His, using t-pHis conjugated to BSA. WB: Human bronchial epithelial cells. IP: pHis proteins from human bronchial epithelial cells.
  • General notes

    It is recommended the antibody is centrifuged at 11200 RCF at 4°C for 10 minutes before use and supernatant used for analysis.

    The following buffers and conditions for the Western blot are suggested:

    Lysis buffer: 150 mM sodium chloride, 0.5 % sodium deoxycholate,  1% Triton X-100, 0.1 % sodium dodecyl sulfate, 10 mM sodium fluoride, 5 mM sodium orthovanadate, 10 mM sodium pyrophosphate, protease inhibitor cocktail, 50 mM tris base, pH 9.0.

    Denature samples in 5x loading buffer with; 10 %  lithium dodecyl sulphate, 40% glycerol, 0.02% bromophenol blue, 50 mM ethylenediaminetetraacetic acid disodium salt dihydrate, 500 mM dithiothreitol, 300 mM tris base, pH 8.8 for 30 min at RT (do not heat).

    Block in; 10 mM tris-hydrochloride, 165 mM sodium chloride, pH 8.0, 0.05 % (v/v), Tween 20, 0.2 % (v/v) freshwater fish gelatin at RT for 1 hr.

    As a control phosphohistidine can be reduced/abolished by reducing the pH (pH 2-4) in denaturing buffer and then heating between 60-90 ºC for > 30 min.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.40
    Preservative: 0.05% Sodium azide
    Constituents: Tris glycine, 0.87% Sodium chloride
  • Concentration information loading...
  • Purity

    Affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG

Applications

Our Abpromise guarantee covers the use of ab231709 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 1/4000.
IP Use at an assay dependent concentration.
Competitive ELISA Use at an assay dependent concentration.

Target

  • Alternative names

    • tele-phosphohistidine antibody

Images

  • Competitive ELISA of ab231709 against phosphorylated amino acids and His, using τ-pHis conjugated to BSA.

  • Anti-phosphohistidine antibody (ab231709) at 1/4000 dilution + Human bronchial epithelial cell lysate at 100 µg
  • Immunoprecipitation of pHis proteins from human bronchial epithelial cells using ab231709. Western blot of immunoprecipitates using ab231709 at 1/4000 dilution.

    pHis antibodies are immobilised using protein G sepharose. Elution refers to batch extraction with competitor. pHis antibody used to probe proteins on PVDF membrane.

    Right pair of images: pHis is known to be acid labile: Acid treatment (pH 7, 60ºC 30 minutes) of lysate before IP.

     

References

ab231709 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

There are currently no Customer reviews or Questions for ab231709.
Please use the links above to contact us or submit feedback about this product.

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Sign up