Embryos for immunofluorescent detection were processed following the protocol proposed by (Ungos et al., 2003). Embryos were fixed in 4 % Paraformaldehyde in PBS (Phosphate buffered saline, Oxoid) overnight at 4 °C. They were washed three times for 5 minutes in PBTriton (0.1 % Triton X-100 Sigma-Aldrich in PBS) and three times for one hour in MilliQ water. Embryos were incubated in block solution (1 % DMSO, 5 % Goat/Sheep serum diluted in PBTriton) for 2 to 3 hours. They were then incubated at room temperature overnight in polyclonal mouse serum primary antibody (Polyclonal IgG Rabbit Anti-GFP Primary Antibody, Invitrogen, A11122) diluted 1:500 in block solution. Embryos were washed in PBTriton once briefly and three times for one hour. They were incubated overnight 4 °C in Alexa Fluor 488 fluorescent anti-rabbit secondary antibody (Polyclonal IgG Alexa Fluor 488 donkey anti-rabbit, Invitrogen, A21206) diluted 1:200 in block solution. Embryos were then washed once briefly and three times for 30 minutes in PBTriton. They were stored in 50 % glycerol for imaging and storage.
Polyclonal igg rabbit anti gfp primary antibody
Manufactured by Thermo Fisher Scientific
The Polyclonal IgG Rabbit Anti-GFP Primary Antibody is a laboratory reagent used for the detection and localization of Green Fluorescent Protein (GFP) in various biological samples. This antibody is produced in rabbits and recognizes the GFP protein, allowing researchers to visualize and study the expression and distribution of GFP in their experiments.
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2 protocols using polyclonal igg rabbit anti gfp primary antibody
1
Immunofluorescent Detection of Embryonic Proteins
2
Immunofluorescent Detection of GFP in Embryos
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Embryos for immunofluorescent detection were processed following the protocol proposed by (Ungos et al., 2003 ). Embryos were fixed in 4% paraformaldehyde in PBS (phosphate‐buffered saline; Oxoid, Basingstoke, Hampshire, UK) overnight at 4°C. They were washed three times for 5 min in PBTriton (0.1% Triton X‐100 Sigma‐Aldrich in PBS) and three times for 1 h in MilliQ water. Embryos were incubated in block solution (1% DMSO, 5% Goat/Sheep serum diluted in PBTriton) for 2–3 h. They were then incubated at room temperature overnight in polyclonal mouse serum primary antibody (polyclonal IgG rabbit anti‐GFP primary antibody; Invitrogen, A11122) diluted 1:500 in block solution. Embryos were washed in PBTriton once briefly and three times for 1 h. They were incubated overnight at 4°C in Alexa Fluor 488 fluorescent anti‐rabbit secondary antibody (polyclonal IgG Alexa Fluor 488 donkey anti‐rabbit; Invitrogen, A21206) diluted 1:200 in block solution. Embryos were then washed once briefly and three times for 30 min in PBTriton. They were stored in 50% glycerol for imaging and storage.
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