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Cytokine ELISA Protocol

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 of 8 votes.

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By vathsala on 06-Nov-2008

Sir,
I need a protocol for IFN gamma measurement from serum by ELISA. I have got a kit from peprotech. I want to know how to dilute serum for the assay. Thanking you.

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By sana on 19-Oct-2008

I need to protocol for IL-2 ,IL-12,INF-gamma measurrment by ELISA.
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By DEVASIER. B, M PHARM on 16-Apr-2008

I NEED THE PROCEDURE IN ANALYSIS OF CYTOKINE USING ELISA
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By Patrisio on 16-Mar-2008

Hi, I am cytokine assays on PBMCs co-incubated with Lactic acid bacteria. very useful information
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By arun on 11-Jun-2007

respected Sir\ Mam,
i am doing my SRF in JIPMER, Pondycherry regarding cytokine assay using ELISA, i will be gratefull if you could please send some protocol and procedure to develop ELISA for above said purpose. thanking you.
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By Gideon on 30-May-2007