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NFκB is regulated by the IκB family of inhibitory proteins. Phosphorylation of IκBα can be triggered by a wide range of extracellular signals, including cytokines and chemokines. It leads, ultimately, to activation of NFκB. Thus, analysis of the phosphorylation state of IκBα provides insights about NFκB and the many genes it regulates. (For additional tools for the study of NFκB, see TransAM™ NFκB p50, p52, p65 and Family Kits and NFκB-related antibodies).
FunctionELISA Kits utilize the Sandwich ELISA technique to capture and quantifiably measure the amount of a specific protein present in a sample. Sandwich ELISAs use two antibodies that recognize different epitopes on the protein being studied. The first antibody is immobilized in the wells of an ELISA plate. When cell lysate is added, the specific protein of interest is bound by this Capture Antibody. A second antibody, called the Detecting Antibody, is then added. It binds to the bound protein. The FunctionELISA IκBα and TRAIL Kits use a horseradish peroxidase (HRP) or alkaline phosphatase (AP) conjugated secondary antibody that binds to the Detecting Antibody to quantify the amount of protein bound by the Capture Antibody (Figure 1A). FunctionELISA Cytochrome c utilizes a biotinylated Detecting Antibody and HRP conjugated to streptavidin for accurate detection of cytochrome c (Figure 1B).
Figure 1: Sandwich ELISA schematics.
Capture and Detecting Antibodies are used for sensitive, accurate quantification of the antigen of interest (Ag).
Why use FunctionELISA?
- Quantitative results in just hours
- ELISA format eliminates need to run, blot and develop gels
- Analyze multiple samples in low-volume, high-throughput experiments
- Complete kit with all required reagents and controls
- Ready-to-use format with capture antibody precoated on the plate
- Ability to assay both cell and tissue samples
