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文献和实验Anti-GFP/BrdU In Situ Immunostaining
by adding 1 mL PBS, 3' each. Add 0.3 mL rabbit anti-GFP (1:200 v/v, plus 1% goat serum in PBS) and incubate 1 hr. at room temp. Wash 3x in PBS. Post-fix in 1 mL 4% paraformaldehyde x15', room temp. Wash 3x in PBS. Denature DNA with 1 mL
-purified goat anti-mouse IgG (Jackson Labs; 1 mg/ml stock used at 1:300 dilution) Texas Red-conjugated affinity-purified donkey anti-rabbit (Jackson Labs;1 mg/ml stock used at 1:200 dilution)
Immunostaining with Anti-Digoxigenin Alkaline Phosphatase
Ususally 1:1000 (v/v) in PBS. Wash in PBS 2' x3. Equilibrate slides in Buffer 1, RT x 5'. Incubate slides in Blocking Buffer, RT x 30' Incubate with anti-Dig-AP conjugate (1:2000 in Blocking buffer.), 2 hrs. RT, or o.n. at 4ºC.
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