Cell death detection in Xenopus embryos by ELISA

Sample preparation

Wash embryos 1 x in 25% MMR

Remove excess buffer

Add 10 volumes "incubation buffer", i.e. 50 µl for 5 embryos

Lyse the embryos by gently pipetting up and down using a large blue tip         Use a negative control, as shearing of genomic DNA may give false positive signals

Dilute 10 µl lysate with 190 µl "incubation buffer"         Consider testing other dilutions

Incubate this sample for 30 min. at 4°C

Centrifuge at 13,000 rpm (eppendorf) for 10 min. at 4°C

Remove 160 µl supernatant / embryo, avoiding pellet contamination of the supernatant

Samples can be stored, if not used the same day, in aliquots at -20°C

Do ELISA according to manufacturer's instructions.