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ELISA Measurement of Mouse IL-2
Published: Jan 5, 2012 DOI: 10.21769/BioProtoc.169 Views: 12087
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Abstract
Interleukin-2 (IL-2) is a cytokine secreted by T cells that is essential for immune system activation. This protocol is routinely used for quantification of IL-2 concentration in the supernant of cultured lymphocytes under various stimulations and co-culturing conditions. Upon slightly modification and optimization, this protocol can also be adapted to quantitatively measure other secreted proteins and bio-molecules.
Materials and Reagents
- BD OptEIA ELISA set including IL-2 standard, capture antibody, detection antibody/enzyme reagent (BD Biosciences, catalog number: 555148 )
- Assay diluent (BD Biosciences, catalog number: 555213 ) or medium made of half RPM1640/10% FCS/PS and half RPM1640
- Substrate solution (BD Biosciences, catalog number: 555214 )
- NaCl
- NaHCO3
- Na2CO3
- Na2HPO4
- KH2PO4
- KCl
- Tween-20
- H2SO4
- Coating buffer (see Recipes)
- Wash buffer (PBST, pH 7.0) (see Recipes)
- Stop solution (see Recipes)
Equipment
- ELISA plates (100 plates/case) (BD Biosciences, catalog number: 353279 ) and plate sealers (100 plates/case) (PGC Scientifics, catalog number: 045-826 )
- 96-well plates for dilution (SARSTEDT AG, catalog number: 82.1583 )
- Multichannel pipette and pipette tips (eBay, catalog number: RT-L200F )
- Reagent reservoir (50 ml, 5/bag) (Corning, Costar®, catalog number: 4870 )
- ELISA micro plate reader
Procedure
Category
Immunology > Antibody analysis > Antibody detection
Biochemistry > Protein > Immunodetection > ELISA
Immunology > Immune cell function > Cytokine
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