6.2.2. Avoiding prophenoloxidase (PPO) activation while collecting haemolymph

Per Kopáček et al. (1995):

  1. Precool the larvae for 15 min at 4ºC.
  2. Collect the haemolymph per Section 6.2.1.
  3. Flush the haemolymph from the capillary into an Eppendorf tube kept on ice.
  4. Add ice cold CA-CAC buffer (20 mM CaCl2 and 10 mM Na-cacodylate, pH 6.5).
  5. Vigorously agitate the tube.
  6. Freeze immediately in liquid nitrogen.
  7. Store the frozen haemolymph at -20ºC.