概述

• 产品名称Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187)抗体[E337]
     

• 描述

  兔单克隆抗体[E337] to Erk1 (pT202/pY204) + Erk2 (pT185/pY187)

• 特异性The antibody detects ERK1 phosphorylated on Threonine 202 and Tyrosine 204 and ERK2 phosphorylated on Threonine 185 and Tyrosine 187.

• 经测试应用WB,IHC-P,Flow Cyt,ICC/IF

• 种属反应性

  与反应: 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).

• 阳性对照    

• WB: Serum starved A431 cell lysate treated with EGF. IHC-P: Human thyroid gland cancer tissue. ICC/IF: A431 cells +- EGF.

• 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.

  Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

性能

• 形式Liquid

• 存放说明Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.

• 存储溶液pH: 7.40
  Preservative: 0.01% Sodium azide
  Constituents: 50% Glycerol, 0.05% BSA

• 
  

• 纯度IgG fraction

• 克隆单克隆

• 克隆编号E337

• 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

• Neuroscience

• Neurology process

• Neurodegenerative disease

• Alzheimer's disease

• Tangles & Tau

• <a javascript:void(0)>Neuroscience

• Neurotransmission

• Intracellular Signaling

• Kinases

Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody [E337] 图像

• 

  Western blot - Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody [E337] (ab32538)          

     Lane 1 : Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody [E337] (ab32538) at 1/500 dilution
  Lane 2 : Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody [E337] (ab32538) at 1/2000 dilution
  Lane 3 : Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody [E337] (ab32538) at 1/10000 dilution
  
  Lane 1 : A431 treated with EGF for 10 minutes
  Lane 2 : A431 treated with EGF for 10 minutes
  Lane 3 : A431 treated with EGF for 10 minutes
  
  Lysates/proteins at 0.1 µg/ml per lane.
  
  Secondary
  Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution
  
  Predicted band size : 42 , 44 kDa
  Observed band size : 42.44 kDa (why is the actual band size different from the predicted?)
  
  
  Exposure time : 3 minutes    

  First-antibody diluted with 1% BSA.

   

   

• 

  Western blot - Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) [E337] antibody (ab32538)          

     
  developed using the ECL technique
  
  Performed under reducing conditions.
  
  Predicted band size : 42 , 44 kDa
  
  
  Exposure time : 16 minutes    

  THP1 cells were incubated at 37°C for 3 minutes with vehicle control (0 μM) and different concentrations of ZK 756326 (ab120814). Increased expression of ERK1 (phospho T202 + Y204 ) + ERK2 (phospho T185 + Y187) (ab32538) in THP1 cells correlates with an increase in ZK 756326 concentration, as described in literature.

  Whole cell lysates were prepared with RIPA buffer (containing protease inhibitors and sodium orthovanadate), 10 μg of each were loaded on the gel and the WB was run under reducing conditions. After transfer the membrane was blocked for an hour using 5% BSA before being incubated with ab32538 at 1/500 dilution and ab17942 at 1 μg /ml overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP (ab97051) at 1/10000 dilution and visualised using ECL development solution.

   

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) [E337] antibody (ab32538)          

     

  Immunohistochemical analysis of paraffin-embedded human thyroid gland cancer using anti-ERK1(pT202/pY204)/ERK2(pT185/pY187) (ab32538) at dilution of 1:50.

   

• 

  Immunocytochemistry/ Immunofluorescence - Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody [E337] (ab32538)          

     

  Immunocytochemistry/Immunofluorescence analysis of 4% paraformaldehyde A431+-EGF(100ng/ml,5min) labelling Erk1 (pT202/pY204) + Erk2 (pT185/pY187) with ab32538 at dilution of 1/200. The secondary antibody used was Alexa Fluor^® 488 Goat-Anti-Rabbit IgG (ab150077) at dilution of 1/400. The counter stain was done with DAPI (blue).

   

• 

  Flow Cytometry - Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) [E337] antibody (ab32538)          

     

  Overlay histogram showing HeLa cells stained with ab32538 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32538, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was a goat anti-rabbit DyLight® 488 (IgG; H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.