製品の概要

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab10321 in the following tested applications.

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

アプリケーション Abreviews 特記事項
WB Use a concentration of 1 µg/ml.
ICC/IF 1/200.
IHC-P Use a concentration of 1 µg/ml.
IHC-FoFr Use a concentration of 1 µg/ml.

ターゲット情報

  • 関連性The phosphorylation of specific tyrosine residues has been shown to be a primary mechanism of signal transduction during normal mitogenesis, cell cycle progression and oncogenic transformation, its role in other areas such as differentiation and gap junction communication, is a matter of active and ongoing research. Antibodies that specifically recognize phosphorylated tyrosine residues have proved to be invaluable to the study of tyrosine phosphorylated proteins and the biochemical pathways in which they function.

Anti-Phosphotyrosine antibody [PY20] 画像

  • 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.
  • 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.
  • 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.
  • 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
    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.

Anti-Phosphotyrosine antibody [PY20] (ab10321) 使用論文

This product has been referenced in:
  • Zalk R  et al. Structure of a mammalian ryanodine receptor. Nature 517:44-9 (2015). Read more (PubMed: 25470061) »
  • Zoppi N  et al. GLUT10 deficiency leads to oxidative stress and non-canonical avß3 integrin-mediated TGFß signalling associated with extracellular matrix disarray in arterial tortuosity syndrome skin fibroblasts. Hum Mol Genet N/A:N/A (2015). WB ; Human . Read more (PubMed: 26376865) »

See all 24 Publications for this product

Product Wall

Application Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample Rat Tissue sections (Brain sections)
Specification Brain sections
Fixative Paraformaldehyde
Antigen retrieval step None
Permeabilization No
Username

Dr. Sophie Pezet

Verified customer

投稿 Jun 12 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample African Green Monkey Cell lysate - whole cell (COS1 cells)
Loading amount 10 µg
Specification COS1 cells
Gel Running Conditions Reduced Denaturing (8%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

投稿 Apr 07 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (HEK 293)
Loading amount 10 µg
Specification HEK 293
Gel Running Conditions Reduced Denaturing (8%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

投稿 Apr 07 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Tissue lysate - whole (Testis)
Loading amount 200 µg
Specification Testis
Gel Running Conditions Reduced Denaturing (10% acrylamide)
Blocking step BSA as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 4°C
Username

Abcam user community

Verified customer

投稿 Oct 28 2008

Application Western blot
Sample Human Cell lysate - whole cell (Human Aortic Endothelial Cell)
Loading amount 30 µg
Specification Human Aortic Endothelial Cell
Treatment 200ng/ml VEGF
Gel Running Conditions Reduced Denaturing (4-20% SDS-PAGE)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

投稿 Aug 08 2008

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"