ERK MAPK Activation Kit

Purchase Details 
 
 
Product Contact 
   Sales Contact
     Service Contact
 
     United States +1 800 874 3723Canada +1 800 874 3723Caribbean +1 800 874 3723Mexico +1 815 968 0747Argentina +1 815 968 0747Chile +1 815 968 0747Other Latin Amer +1 815 968 0747Austria +32 5 385 71 84Belgium +32 5 385 71 84Denmark +32 5 385 71 84Finland +32 5 385 71 84France +33 800 508 215Germany +49 2289 125 650Ireland +32 53 85 71 84Italy +32 5 385 71 84Luxembourg +32 5 385 71 84Netherlands +31 76 503 1880Norway +32 5 385 71 84Portugal +32 5 385 71 84Russia +32 5 385 71 84Spain +32 5 385 71 84Sweden +32 5 385 71 84Switzerland +0800 56 31 40United Kingdom +44 800 252 185Other Europe +32 5 385 71 84China +86 10 80499033India +1 815 968 0747Japan +81 45 453 9229Middle East +1 815 968 0747Other Asia +1 815 968 0747South Africa +1 815 968 0747Other Africa +1 815 968 0747Australia +1 815 968 0747Oceania +1 815 968 0747 
 
 
 
 
 
 
 

ORDER NOW - ERK MAPK Activation Kit
 
 
 ERK MAPK Activation Kit
 Product# Price  Description  Pack Size  
         
 K0100071      -    ERK MAPK (green) Activation   5 x 96   SELECT   
 R0105091      -    ERK MAPK (green) Activation  50 x 96   SELECT   
 Details All Product Numbers 
  Details All Product Numbers
 

Figure 3. Dose response curve for PMA in NIH 3T3 cells.  NIH 3T3 cells were stimulated with PMA for 30 minutes, then labeled as per the ERK MAPK Activation kit protocol.  The dose response curve was used to determine an EC50 of 2.7 ng/mL.

Figure 1. Images of fluorescently lable NIH 3T3 cells before (left) and after (right) activation of ERK by PMA (100 ng/mL for 30 min.) Cells were prepared according to the kit protocol.  Nuclei appear blue (Hoechst) and ERK appears green (Alexa Fluor 488). Images were aquired by the ArrayScan HCS Reader.

The ERK MAPK Activation Kit enables ERK quantitation by its translocation to the nucleus. This kit in combination with the ArrayScan HCS Reader and the Cytoplasm to Nucleus Translocation BioApplication software allows automated plate handling, focusing, cell image acquisition and analysis of ERK MAPK activation. For a more detailed description of the image processing algorithm, see the Cytoplasm to Nucleus Translocation BioApplication Guide that accompanies the Cytoplasm to Nucleus Translocation BioApplication software.

Mitogen-activated protein kinase (MAPK) pathways regulate an extensive range of cellular processes including gene transcription, cytoskeletal organization, metabolic homeostasis, cell growth and apoptosis. MAPKs are activated by concomitant Tyr and Thr phosphorylation via a complex multi-step signal transduction cascade. The extracellular signal-regulated kinases (ERK) 1 and 2, also known as p44 and p42 MAPK, accumulate in the nucleus after stimulation with mitogens. Thus the MAPK/ERK cascade constitutes a functional signaling unit that links surface receptor-mediated signals to nuclear events.

 
ERK MAPK Activation Kit Features
A more biologically relevant measure of kinase activation relative to Western blots or in vitro kinase assays
No cell lysis, purification, or filtration steps
Immunofluorescence based assay provides sensitive quantitative and qualitative results without radioactivity
Kit antibody preferentially binds to dually phosphorylated (fully active) ERK, with minimized cross-reactivity to other MAPKs
Manual sample preparation/processing requires approximately 3 hours post-compound treatment
The ERK MAPK Activation Kit provides the reagents and protocol necessary to quantify ERK activation by directly measuring its translocation from the cytoplasm to the nucleus. This fixed end-point assay is performed on live cells growing on standard high-density microplates. Inhibitors of ERK translocation are screened by stimulating cells with a control inducer, such as phorbol 12-myristate 13-acetate (PMA) after exposing cells to the test compounds. The kit is supplied with an anti-ERK primary antibody and a DyLight 488-conjugated secondary antibody. The nuclear region is identified by the nuclear dye, Hoechst, also included in the kit.

   Product Detail 

 
The HCS Reagent Kit for ERK Activation provides high-content screening (HCS)-quality fluorescence reagents and a validated protocol for the preparation and processing of samples for the measurement of ERK kinase activation.