• Compound/Extract Preparation
  • Treatment
  • NO measurement (Next day) and adding MTT solution

Compound/Extract Preparation

  • Extract = 16.42 mg/tube
  • Make stock 100 mg/ml –> add 0.1642 ml = 164 ul

Treatment

  • Compound/Extract pre-treatment 50 ul for 30 min
  • LPS treatment 50 ul for 24 h

NO measurement (Next day) and adding MTT solution

  • Move supernatant (100 ul) to a new 96 well plate
  • Mix Griess reagent A and B (1:1) -> approximate volume needed
  • Add mixed Griess reagent (100 ul/well) into 96 well plate containing supernatant
  • Measure the absorbance (540 nm)
  • Calculate the NO concentration according to NO standard curve