Rabbit anti gfp

Primary antibodies: rat anti-Src1 (this work), rabbit anti-NE81 [18 (link)], rat YL1/2 [32 (link)], rabbit anti-GFP [33 (link)], rabbit anti-GFP (Molecular Probes, A-6455; Life Technologies, Carlsbad, CA, USA). AlexaFluor conjugated secondary antibodies and streptavidin-AlexaFluor 488 were purchased from Life Technologies (Carlsbad, CA, USA), streptavidin-CIP (calf intestine alkaline phosphatase) and anti-rabbit-CIP/anti-rat-CIP from Sigma-Aldrich (Deisenhofen, Germany).

Briefly, Tg(cmyb:EGFP) embryos were incubated with 10 mM BrdU (Sigma) for 30 minutes and incubated with egg water for another 2 hours, then fixed in 4% paraformaldehyde (PFA) at 3 dpf. After dehydration and rehydration, the embryos were treated with Proteinase K (10 mg/ml, Finnzyme) for 30 minutes and re-fixed in 4% PFA for 30 minutes. After blocking with blocking buffer (2 mg/ml BSA, 10% FBS, 0.3% Triton-X100 and 1% DMSO in PBST), the embryos were stained with rabbit anti-GFP (Invitrogen) primary antibody and Alexa Fluor 488-conjugated anti-rabbit (Invitrogen) secondary antibodies. The embryos were fixed again, treated with Proteinase K for the second time, and re-fixed in 4% PFA. The embryos were then incubated with 2 N HCl for 1 hour and stained with mouse anti-BrdU (Roche) and rabbit anti-GFP (Invitrogen) antibodies. Finally, Alexa Fluor 594-conjugated anti-mouse and Alexa Fluor 488-conjugated anti-rabbit (Invitrogen) secondary antibodies were used. Images were taken using Olympus FV 1000 confocal microscopy equipped with the FV10-ASW version3 software.
Terminal transferase UTP nick end labeling (TUNEL) was performed using the In Situ Cell Death Detection Kit, TMR red (Roche) according to the manufacturer's recommendations.