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AlphaScreen SureFire IKKa (p-Ser176/Ser180) Assay Kit - 50,000 Points

The AlphaScreen® SureFire® p-IKKα (Ser176/180) assay is a sandwich immunoassay for quantitative detection of phospho-IKKα (phosphorylated on Ser176/180) in cellular lysates using Alpha Technology. Please note that an AlphaScreen Protein A kit, which contains the Alpha Beads necessary to run this assay, needs to be purchased separately in addition to the AlphaScreen SureFire kit.

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TGRKAS500
500 assay points
1653.00 USD
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TGRKAS10K
10,000 assay points
7200.00 USD
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TGRKAS50K
50,000 assay points
17700.00 USD
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Detail Information

Alpha SureFire® kits can be used for:

  • Cellular kinase assays
  • Receptor activation studies
  • Screening
 

Formats:

  • The HV 96-well kits contain enough reagents to run 100 wells in 96-well format, using a 50 µL reaction volume.
  • The 500 point kit contains enough reagents to run 500 wells in 384-well format, using an 11 µL reaction volume.
  • The 10,000 point kit contains reagents to run 10,000 wells in 384-well format, using an 11 µL reaction volume.
  • The 50,000 point kit contains enough reagents to run 50,000 wells in 384-well format, using an 11 µL reaction volume.
 

In the AlphaScreen® SureFire® assay, Donor beads are coated with streptavidin to capture one of the antibodies, which is biotinylated. Acceptor beads are coated with Protein A to immobilize the other antibody. In the presence of phosphorylated protein, the two antibodies bring the Donor and Acceptor beads close together, generating signal. The amount of light emission is directly proportional to the amount of phosphoprotein present in the sample.

Specifications

Assay Pathway NFκB
Assay Target IKKα
Assay Target Class Phosphoprotein
Automation Compatible Yes
Detection Method Alpha
Experimental Type In vitro
Molecular Modification Phosphorylation
Product Brand Name AlphaScreen SureFire
Second Messenger Release Protein phosphorylation
Shipping Condition Blue Ice
Target Species Human
Unit Size 50,000 assay points
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Application Note

A Practical Approach to Cell Signaling Pathway Analysis

Cellular kinase signal transduction pathways are involved in the regulation of many important cellular processes suchas cell survival, differentiation and apoptosis. Kinase signaling networks are typically characterized by multiple kinases arranged in cascades containing nodes with feedback loops, and crosstalk between pathways.

PDF 1 MB
Quick Guide to AlphaScreen SureFire Assay Optimization

Screening kinase activities in cell-based assays offers advantages over the more traditional biochemical approach of using a purified recombinant enzyme to phosphorylate a substrate, since a cell-based assay gives information on a compound’s activity in a more biological context. However, getting initial results with the AlphaScreen® SureFire® assay is highly dependent on optimal cell culture conditions, and often requires that multiple parametersbe optimized in the first set of experiments.

PDF 307 KB

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Technical Note

Cell lines validated for use with AlphaScreen® SureFire® kits

Cell lines validated for use with AlphaScreen® SureFire® kits

PDF 55 KB