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AlphaScreen SureFire Total NFkB p65 Assay Kit - 10,000 Points

The AlphaScreen® SureFire® total NFκB assay is a sandwich immunoassay for quantitative detection of NFκB (both phosphorylated and non-phosphorylated) in cellular lysates using Alpha Technology. This assay is intended to be used as a normalization for phosphorylation studies. 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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Unit Size
List Price
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500 assay points
1615.00 USD
10,000 assay points
7100.00 USD
50,000 assay points
17400.00 USD
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Detail Information

Alpha SureFire® kits can be used for:

  • Cellular kinase assays
  • Receptor activation studies
  • Screening


  • 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 target 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 protein present in the sample.


Assay Pathway NFκB
Assay Target NFκB
Assay Target Class Protein
Automation Compatible Yes
Detection Method Alpha
Experimental Type In vitro
Product Brand Name AlphaScreen SureFire
Quantity in a Package Amount 10000.0 Units
Shipping Condition Blue Ice
Target Species Human
Unit Size 10,000 assay points
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Application Note

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.

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Protocol for AlphaScreen® SureFire® Total NFkB Assay Kits

General Information on the AlphaScreen® SureFire® Total NF-?B p65 assay,The AlphaScreen® SureFire® Total NF-?B p65 assay is used to measure endogenous NF-?B p65 in cellular lysates. The assay is an ideal system for the screening of both modulators of receptor activation (e.g. agonists and antagonists) as well as agents acting intracellularly, such as small molecule inhibitors of upstream events.

PDF 391 KB

Technical Note

Cell lines validated for use with AlphaScreen® SureFire® kits

Cell lines validated for use with AlphaScreen® SureFire® kits