Blocking Solution for Xenopus Embryos
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1. Mix 4.6 g maleic acid and 3.5 g NaCl in 200 mL of sterile milliQ H2O in a 1000-mL beaker. Adjust the pH to 7.5 with 10 n NaOH.
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2. Add 8 g of Blocking Reagent (Sigma-Aldrich, 11096176001). Heat the solution in a microwave using 30-sec pulses until the Blocking Reagent is fully dissolved. (The solution should appear milky.)
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3. Cool on ice. Adjust the volume to 380 mL with sterile milliQ H2O.
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4. Add 20 mL of heat-inactivated lamb serum (ThermoFisher, 16070096). (Inactivate the serum by heating in a water bath at 55°C for 30 min, swirling every 5–10 min. Store at −20°C in 20-mL aliquots.)
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5. Store the blocking solution at −20°C in 20-mL aliquots.
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The final concentrations of the reagents are 100 mm maleic acid, 150 mm NaCl, 5% lamb serum, 2% (w/v) blocking reagent.
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