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FunctionELISA™ Cytochrome c

sandwich ELISA for accurate quantification of cytochrome c

Because cytochrome c plays a major role in apoptosis, its measurement can lead to greater understanding of certain diseases on a cellular level. Cytochrome c is an essential component of the mitochondrial respiratory chain. It is a soluble protein that is localized in the intermembrane space and loosely attached to the surface of the inner mitochondrial membrane. Cytochrome c is important to the process of creating cellular energy, the main function of mitochondria. When mitochondria are damaged, cytochrome c is released into the main body of the cell and, if the cell itself is damaged, into the surrounding tissue. The release of cytochrome c is part of the cascade of cellular events that leads to apoptosis, or programmed cell death. When the cycle that leads to apoptosis begins, cytochrome c appears outside the mitochondria within one hour.

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

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FunctionELISA™ Cytochrome c

sandwich ELISA for accurate quantification of cytochrome c

Figure 1: Sandwich ELISA schematics.

Why use FunctionELISA?

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