Antibody staining for cyphonautes larvae
Execute the protocol for antibody staining with Chema to test in cyphonautes larvae of Membranipora. Chosen antibody is phospho-histone which binds to histone in chromatin revealing dividing cells.
26/02/2012
Day one of the standard protocol. Incubating overnight with rabbit primary antibody diluted 1:2500 in PTx+NGS.
27/02/2012
Day two, washes and incubation with secondary (rabbit) antibody (orange-red AlexaFluor 594) diluted 1:200 in PTx+NGS.
28/02/2012
Day three, regular washes and addition of Phallacidin with 1:1000 DAPI. Staining for 1h and mounting in glycerol.
Mounted 3 slides. 1 is good, 2 has most embryos and most dirt, 3 has only a few embryos. Checked the slides in the fluorescent lamp: DAPI and Phallacidin look great and strong; Phospho-histone signal is widespread in the whole embryo, but seems to be slightly stronger in the apical region.