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ELISA Measurement of Mouse IL-2

GH Guo N. Huang

Published: Jan 5, 2012 DOI: 10.21769/BioProtoc.169 Views: 12060

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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

  1. BD OptEIA ELISA set including IL-2 standard, capture antibody, detection antibody/enzyme reagent (BD Biosciences, catalog number: 555148 )
  2. Assay diluent (BD Biosciences, catalog number: 555213 ) or medium made of half RPM1640/10% FCS/PS and half RPM1640
  3. Substrate solution (BD Biosciences, catalog number: 555214 )
  4. NaCl
  5. NaHCO3
  6. Na2CO3
  7. Na2HPO4
  8. KH2PO4
  9. KCl
  10. Tween-20
  11. H2SO4
  12. Coating buffer (see Recipes)
  13. Wash buffer (PBST, pH 7.0) (see Recipes)
  14. Stop solution (see Recipes)

Equipment

  1. ELISA plates (100 plates/case) (BD Biosciences, catalog number: 353279 ) and plate sealers (100 plates/case) (PGC Scientifics, catalog number: 045-826 )
  2. 96-well plates for dilution (SARSTEDT AG, catalog number: 82.1583 )
  3. Multichannel pipette and pipette tips (eBay, catalog number: RT-L200F )
  4. Reagent reservoir (50 ml, 5/bag) (Corning, Costar®, catalog number: 4870 )
  5. ELISA micro plate reader

Procedure

Copyright: © 2012 The Authors; exclusive licensee Bio-protocol LLC.

Category

Immunology > Antibody analysis > Antibody detection

Biochemistry > Protein > Immunodetection

Immunology > Immune cell function > Cytokine

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