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Anti-Phosphotyrosine antibody [PY20] (ab10321)

Key features and details

  • Mouse monoclonal [PY20] to Phosphotyrosine
  • Suitable for: WB, ICC/IF, IHC-P, IHC-FoFr
  • Reacts with: Species independent
  • Isotype: IgG2b

Overview

  • Product name

    Anti-Phosphotyrosine antibody [PY20]
    See all Phosphotyrosine primary antibodies

  • Description

    Mouse monoclonal [PY20] to Phosphotyrosine

  • Host species

    Mouse

  • Tested applications

    Suitable for: WB, ICC/IF, IHC-P, IHC-FoFrmore details

  • Species reactivity

    Reacts with: Species independent

  • Immunogen

    Chemical/ Small Molecule corresponding to Phosphotyrosine conjugated to keyhole limpet haemocyanin.

  • General notes

    This antibody clone is manufactured by Abcam.

    This is a standard clone used to detect phosphotyrosine.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

    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

Images

  • Western blot - Anti-Phosphotyrosine antibody [PY20] (ab10321)
    Western blot - Anti-Phosphotyrosine antibody [PY20] (ab10321)
    All lanes : Anti-Phosphotyrosine antibody [PY20] (ab10321) at 1 µg/ml

    Lane 1 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lane 2 : NIH 3T3 treated with Vanadate and PDGF Whole Cell Lysate

    Lysates/proteins at 5 µg per lane.

    Secondary
    All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Exposure time: 1 minute


    Cells were serum starved overnight and then incubated at room temperature for 10mins in a final concentration of 1mM sodium vanadate. PDGF was then added at a final concentration of 5ng/ml and cells were incubated at 37ºC for 30mins. Vanadate inhibits endogenous phosphatases and PDGF stimulates phosphorylation. Western blots of NIH 3T3 cell lysates treated with vanadate and PDGF show an array of phosphorylated tyrosine compared to controls.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Phosphotyrosine antibody [PY20] (ab10321)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Phosphotyrosine antibody [PY20] (ab10321) This image is courtesy of Sophie Pezet, Univ London Kings Coll, United Kingdom
    Immunofluorescent staining with ab10321 mouse monoclonal [PY20] phosphotyrosine antibody in the rat cortex. Cells stained appear to be microglial cells. Picture taken with objective X20. Protocol: IHC free-floating protocol using 4%PFA fixed brain tissue. Rats were intracardially perfused with 4% PFA. Tissue was post-fixed overnight in the same fixative, cryoprotected in 20% sucrose and frozen in OCT. Primary antibody ab10321 was used at 1ug/ml incubated overnight at room temperature. Secondary antibody was Alexa Fluor 488 used at 1/1000, 2h incubation at room temperature. Image recoloured in Adobe photoshop.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Phosphotyrosine antibody [PY20] (ab10321)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Phosphotyrosine antibody [PY20] (ab10321) This image is courtesy of Sophie Pezet, Univ London Kings Coll, United Kingdom
    Immunofluorescent staining with ab10321 mouse monoclonal [PY20] phosphotyrosine antibody in the rat spinal cord. Cells stained appear to be microglial cells. Picture taken with  X40 objective. Protocol: IHC free-floating protocol using 4% PFA fixed spinal cord tissue. Rats were intracardially perfused with 4% PFA. Tissue was post-fixed overnight in the same fixative, cryoprotected in 20% sucrose and frozen in OCT. Primary antibody ab10321 was used at 1ug/ml incubated overnight at room temperature. Secondary antibody was Alexa Fluor 488 used at 1/1000, 2h incubation at room temperature. Image recoloured in Adobe photoshop.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Phosphotyrosine antibody [PY20] (ab10321)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Phosphotyrosine antibody [PY20] (ab10321)
    IHC image of ab10321 staining in Human Normal Hippocampus formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab10321, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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