概述

• 产品名称Anti-Phosphotyrosine抗体[EPR16871]
     

• 描述

  兔单克隆抗体[EPR16871] to Phosphotyrosine

• 经测试应用WB,ICC/IF,IP,Flow Cyt,ELISA

• 种属反应性

  与反应: Mouse, Rat, Human    
   

• 免疫原

  Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Phosphotyrosine.

• 阳性对照    

• WB: Whole cell lysate from A431, L6 and NIH/3T3 cells treated with pervanadate. ICC/IF: C2C12 cells treated with Hydrogen peroxide. Flow Cyt: A431 cells treated with pervanadate. IP: Whole cell extract from A431 cells treated with pervanadate.

• 常规说明

  This product is a recombinant rabbit monoclonal antibody.

  Produced using Abcam’s RabMAb^® technology. RabMAb^® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

性能

• 形式Liquid

• 存放说明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.

• 存储溶液Preservative: 0.01% Sodium azide
  Constituents: 59% PBS, 40% Glycerol, 0.05% BSA

• 
  

• 纯度Protein A purified

• 克隆单克隆

• 克隆编号EPR16871

• 同种型IgG

• 研究领域

• Metabolism

• Pathways and Processes

• Metabolic signaling pathways

• Amino acid metabolism

• Signal Transduction

• Protein Phosphorylation

• Tyrosine Phosphatases

• Signal Transduction

• Metabolism

• Amino Acids

• <a javascript:void(0)>Neuroscience

• Neurotransmission

• Intracellular Signaling

• Phosphatases

Anti-Phosphotyrosine antibody [EPR16871] 图像

• 

  Immunocytochemistry/ Immunofluorescence - Anti-Phosphotyrosine antibody [EPR16871] (ab179530)          

     

  Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized C2C12 (Mouse myoblast cell line) cells labeling Phosphotyrosine with ab179530 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/400 dilution (green). Cytoplasm staining on C2C12 cells is observed. The expression increased after treatment with H2O2 (2mM) for 10 minutes. The nuclear counterstain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
  The negative controls are as follows:-
  -ve control 1: - ab179530 at 1/100 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
  -ve control 2: - ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/400 dilution.

   

   

• 

  Dot Blot - Anti-Phosphotyrosine antibody [EPR16871] (ab179530)          

     

  Dot blot analysis of INSR/IGF-1R (pY1009) phospho peptide (lane 1) and INSR/IGF-1R non-phospho peptide (lane 2) labelling Phosphotyrosine with ab179530 at a dilution of 1/1000. A peroxidase-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/2500).

  Blocking and dilution buffer: 5% NFDM/TBST.

  Exposure time: 3 minutes.

   

• 

  Immunoprecipitation - Anti-Phosphotyrosine antibody [EPR16871] (ab179530)          

     

  Phosphotyrosine was immunoprecipitated from 1mg of A431 (Human epidermoid carcinoma) whole cell extract treated with 1mM pervanadate for 30 minutes with ab179530 at 1/100 dilution. Western blot was performed from the immunoprecipitate using ab179530 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution. Lane 1: A431 treated with 1mM pervanadate for 30 minutes whole cell extract. Lane 2: PBS instead of A431 whole cell extract.

  Blocking and dilution buffer and concentration: 5% NFDM/TBST.
  Multiple bands represent phosph-tyrosine containing proteins precipitated and detected by ab179530.

   

• 

  Western blot - Anti-Phosphotyrosine antibody [EPR16871] (ab179530)          

     All lanes : Anti-Phosphotyrosine antibody [EPR16871] (ab179530) at 1/1000 dilution
  
  Lane 1 : Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysates treated with 1mM pervanadate for 20minutes
  Lane 2 : Untreated Jurkat whole cell lysates
  
  Lysates/proteins at 10 µg per lane.
  
  Secondary
  Goat Anti-Rabbit IgG (H+L) Peroxidase conjugated at 1/1000 dilution    

  Multiple bands represent phosph-tyrosine containing proteins detected by ab179530

   

• 

  Western blot - Anti-Phosphotyrosine antibody [EPR16871] (ab179530)          

     All lanes : Anti-Phosphotyrosine antibody [EPR16871] (ab179530) at 1/10000 dilution
  
  Lane 1 : A431 (Human epidermoid carcinoma) whole cell lysates treated with 50mM pervanadate for 30 minutes
  Lane 2 : Untreated A431 whole cell lysates
  
  Lysates/proteins at 10 µg per lane.
  
  Secondary
  Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution    

  Multiple bands represent phosph-tyrosine containing proteins detected by ab179530.

  Blocking/Dilution buffer: 5% NFDM/TBST.

   

   

• 

  Western blot - Anti-Phosphotyrosine antibody [EPR16871] (ab179530)          

     All lanes : Anti-Phosphotyrosine antibody [EPR16871] (ab179530) at 1/1000 dilution
  
  Lane 1 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysates treated with 1mM pervanadate for 10 minutes
  Lane 2 : Untreated NIH/3T3 whole cell lysates
  
  Lysates/proteins at 10 µg per lane.
  
  Secondary
  Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution    

  Multiple bands represent phosph-tyrosine containing proteins detected by ab179530.

  Blocking/Dilution buffer: 5% NFDM/TBST.

   

• 

  Western blot - Anti-Phosphotyrosine antibody [EPR16871] (ab179530)          

     All lanes : Anti-Phosphotyrosine antibody [EPR16871] (ab179530) at 1/1000 dilution
  
  Lane 1 : L6 (Rat skeletal muscle cell line) whole cell lysates treated with 1mM pervanadate for 20 minutes
  Lane 2 : Untreated L6 whole cell lysates
  
  Lysates/proteins at 10 µg per lane.
  
  Secondary
  Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution    

  Multiple bands represent phosph-tyrosine containing proteins detected by ab179530.

  Blocking/Dilution buffer: 5% NFDM/TBST.

   

• 

  Flow Cytometry - Anti-Phosphotyrosine antibody [EPR16871] (ab179530)          

     

  Flow cytometric analysis of 2% paraformaldehyde-fixed Pervanadate (50mM, 30min.) treated (orange)/untreated (red) A431 (Human epidermoid carcinoma) cells labeling Phosphotyrosine with ab179530 at 1/160 dilution compared with a rabbit monoclonal IgG control (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.

   

• 

  ELISA - Anti-Phosphotyrosine antibody [EPR16871] (ab179530)          

     

  ELISA analysis of various antigens (1 µg/ml) using ab179530 at 1/6400 dilution followed by Alkaline Phosphatase-conjugated Goat Anti-Rabbit IgG (H+L) at 1/2500 dilution.

  S/N = signal-to-noise ratio of phospho- versus nonphospho-peptides.