概述

• 产品名称Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187)抗体[EP197Y]
     

• 描述

  兔单克隆抗体[EP197Y] to Erk1 (pT202/pY204) + Erk2 (pT185/pY187)

• 特异性This antibody detects Erk1 phosphorylated at Threonine 202 and Tyrosine 204 and Erk2 phosphorylated at Threonine 185 and Tyrosine 187.

• 经测试应用WB,IP

• 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 Human Erk1 (pT202/pY204) + Erk2 (pT185/pY187). Swissprot Numbers: P27361, P28482.

• 阳性对照    

• WB: A431 (treated with EGF), SH-SY5Y (treated with NGF), PC-12 (treated with NGF), HeLa and NIH/3T3 whole cell lysates. IP: A431 (treated with EGF) and PC-12 (treated with NGF) whole cell lysates.

• 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. Avoid freeze / thaw cycle.

• 存储溶液pH: 7.20
  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EP197Y

• 同种型IgG

• 研究领域

• Cell Biology

• Other Antibodies

• Oxidative Stress

• Signal Transduction

• Protein Trafficking

• Golgi Proteins

• Stem Cells

• Signaling Pathways

• TGF beta

• Cytoplasmic

• Signal Transduction

• Protein Trafficking

• Vesicle Transport

• Regulation

• Signal Transduction

• Protein Phosphorylation

• Ser / Thr Kinases

• MAPK Pathway

• Signal Transduction

• Protein Phosphorylation

• Tyrosine Phosphatases

• Signal Transduction

• Cytoskeleton / ECM

• Extracellular Matrix

• Structures

• Focal Adhesions

• Immunology

• Innate Immunity

• Cytokines

• Interleukins

• <a javascript:void(0)>Neuroscience

• Neurology process

• Neurodegenerative disease

• Alzheimer's disease

• Tangles & Tau

Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody [EP197Y] 图像

• 

  Dot Blot - Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody [EP197Y] (ab76299)          

     

  Dot blot analysis of single phospho peptide pT202 (Lane 1), single phospho peptide pY204 (Lane 2), souble phospho peptide pT202/pY204 (Lane 3) and non-phospho peptide (Lane 4) labelling Erk1 (pT202/pY204) + Erk2 (p185/pY187) with ab76299 at a dilution of 1/1000. A peroxidase-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody at a dilution of 1/1000.

  Blocking and dilution buffer: 5% NFDM/TBST.

  Exposure time: 3 minutes.

   

• 

  Western blot - Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody [EP197Y] (ab76299)          

     All lanes : Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody [EP197Y] (ab76299) at 1/5000 dilution
  
  Lane 1 : A431 whole cell lysate - untreated
  Lane 2 : A431 whole cell lysate - treated with 100ng/ml EGF for 10 minutes
  
  Lysates/proteins at 10 µg per lane.
  
  Secondary
  Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
  
  Predicted band size : 43, 41 kDa
  Observed band size : 44 kDa (why is the actual band size different from the predicted?)
  
  
  Exposure time : 3 minutes    

  Blocking and dilution buffer: 5% NFDM/TBST.

   

• 

  Western blot - Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody [EP197Y] (ab76299)          

     All lanes : Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody [EP197Y] (ab76299) at 1/10000 dilution
  
  Lane 1 : SH-SY5Y whole cell lysate - untreated
  Lane 2 : SH-SY5Y whole cell lysate - treated with 100ng/ml NGF for 10 minutes
  
  Lysates/proteins at 10 µg per lane.
  
  Secondary
  Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
  
  Predicted band size : 43, 41 kDa
  Observed band size : 42,44 kDa (why is the actual band size different from the predicted?)
  
  
  Exposure time : 15 seconds    

  Blocking and dilution buffer: 5% NFDM/TBST.

   

• 

  Western blot - Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody [EP197Y] (ab76299)          

     All lanes : Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody [EP197Y] (ab76299) at 1/10000 dilution
  
  Lane 1 : PC-12 whole cell lysate - untreated
  Lane 2 : PC-12 whole cell lysate - treated with 100ng/ml NGF for 10 minutes
  
  Lysates/proteins at 10 µg per lane.
  
  Secondary
  Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
  
  Predicted band size : 43, 41 kDa
  Observed band size : 42,44 kDa (why is the actual band size different from the predicted?)
  
  
  Exposure time : 15 seconds    

  Blocking and dilution buffer: 5% NFDM/TBST.

   

• 

  Western blot - Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody [EP197Y] (ab76299)          

     All lanes : Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody [EP197Y] (ab76299) at 1/10000 dilution
  
  Lane 1 : HeLa whole cell lysate - untreated
  Lane 2 : HeLa whole cell lysate - treated with Lambda Phosphatase
  
  Lysates/proteins at 10 µg per lane.
  
  Secondary
  Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
  
  Predicted band size : 43, 41 kDa
  Observed band size : 42,44 kDa (why is the actual band size different from the predicted?)
  
  
  Exposure time : 30 seconds    

  Blocking and dilution buffer: 5% NFDM/TBST.

   

• 

  Western blot - Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody [EP197Y] (ab76299)          

     Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody [EP197Y] (ab76299) at 1/10000 dilution + NIH/3T3 whole cell lysate at 10 µg
  
  Secondary
  Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
  
  Predicted band size : 43, 41 kDa
  Observed band size : 42,44 kDa (why is the actual band size different from the predicted?)
  
  
  Exposure time : 3 minutes    

  Blocking and dilution buffer: 5% NFDM/TBST.

   

• 

  Immunoprecipitation - Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody [EP197Y] (ab76299)          

     

  ab76299 at 1/80 immunoprecipitating Erk1 (pT202/pY204) + Erk2 (pT185/pY187) in A431 whole cell lysate.

  Lane 1 (input): A431 whole cell lysate - treated with 100ng/ml EGF for 10 min (10µg).

  Lane 2 (+): ab76299 + A431 whole cell lysate - treated with 100ng/ml EGF for 10 min.

  Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab76299 in A431 whole cell lysate - treated with 100ng/ml EGF for 10 min.

  For western blotting, ab76299 was used at a dilution of 1/1000 and a HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG, was used as the secondary antibody (1/1500).

  Blocking and dilution buffer: 5% NFDM/TBST.

   

• 

  Immunoprecipitation - Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody [EP197Y] (ab76299)          

     

  ab76299 at 1/80 immunoprecipitating Erk1 (pT202/pY204) + Erk2 (pT185/pY187) in PC-12 whole cell lysate.

  Lane 1 (input): PC-12 whole cell lysate - treated with 100ng/ml NGF for 10 min (10µg).

  Lane 2 (+): ab76299 + PC-12 whole cell lysate - treated with 100ng/ml NGF for 10 min.

  Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab76299 in PC-12 whole cell lysate - treated with 100ng/ml NGF for 10 min.

  For western blotting, ab76299 was used at a dilution of 1/1000 and a HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG, was used as the secondary antibody (1/1500).

  Blocking and dilution buffer: 5% NFDM/TBST.

   

• 

  ELISA - Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody [EP197Y] (ab76299)          

     

  ELISA analysis of various phospho and non-phospho peptides (0-250 ng/ml) labelling Erk1 (pT202/pY204) + Erk2 (pT185/pY187) with ab76299 at a dilution of 1/1000. An Alkaline Phosphatase-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/2500).

  ab76299 has stronger affinity for the double phospho peptide Y204/Y187 than to single phospho peptides T202 or Y204.