赛默飞世尔Thermo Fisher ANTI-GUCY2C

Wholemount staining of embryos

) at room temperature for 15 to 30''. Use nutator (Clay Adams, Fisher cat#14-062) Remove blocking solution and add primary antibody (500 ul), incubated overnight at 4 degC if sensitivity is not a problem. Antibody is added either as a cell culture

Purification of mAb (IgG)

preparation (serum, tissue culture supernatant, or ascites). Apply Binding buffer again ~ 200 ml. Apply Elution buffer ~ 100 ml. Collect 3 ml fractions in 5 ml tubes with 700 ul 1 M Tris pH 9.0 already in bottom of tube to neutralize

Single and Double FISH protocols for Drosophila

mM dithiothreitol, 20 mM spermidine), 1ul RNAse inhibitor (1unit/ul) and sterile, RNAse-free water to make the final reaction volume equal to 18 ul. Add 2 ul of the appropriate RNA polymerase (T7 or T3), mix well and incubate at 37oC for 2 hours.