Cytokine ELISA Protocol

Cytokine elisa protocol:

This protocol described in detail the sandwich ELISA (enzyme-based
immunoassay) method for detection of soluble cytokines and chemokines
in tissue lysate, serum or other fluids.

General cytokine ELISA procedure includes:

- Capture antibody: add diluted high specificity antibody in 96 well
ELISA plate overnight.
- Blocking non-specific binding site of the capture antibody:
- Add standards and samples for overnight incubation and wash.
- Detect by biotinylated anti-cytokine detection antibody followed by
avidin-horseradish peroxidase (Av-HRP).
- Add substrate (ABTS Substrate) and read optical density (O.D. value)
with microplate reader at wavelength 405nm.

Commercial available cytokine ELISA kits normally detect following
soluble cytokines:
proinflammatory cytokines: GM-CSF, IL-1?, IL-1RA, IL-6, IL-8, and TNF-?;
Th1/Th2 distinguishing cytokines: IFN-?, IL-2, IL-2R, IL-4, IL-5, and IL-10
nonspecific acting cytokines: IFN-?, IL-7, IL-12p40/p70, IL-13, IL-15, and IL-17
chemokines: eotaxin, IP-10, MCP-1, MIP-1?, MIP-1?, MIG, and RANTES.

Reference:
Cytokine ELISA protocol

Last update 09-Jan-2005, Rating Very Good of 6 votes.