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ELISpot Protocol

Detailed procedure for ELISpot. View our ELISpot Troubleshooting to solve your problem.

Reagents

General ELISpot Procedure

Typical Stimulates

  1. LPS to stimulate IL1β, IL6 secretion
  2. PMA and ionomycin stimulate IL2, IL4 secretion
  3. PHA, 10 µg/ml for IFN gamma
  4. Anti-CD3/CD28 antibodies for IFN gamma, IL4, IL10 and Granzyme B

Indirect ELISpot

If the cells take some time to respond to stimulation, they may require pretreatment with the stimulant in a separate 96-well culture dish before transferring to the ELISpot plate.

Control Setting

Precautions