Blocking Buffer for TEM
MATERIALS
Reagents
Phosphate-buffered saline (PBS)
Bovine serum albumin (BSA) (Sigma A9647)
Cold water fish skin gelatin (Sigma G7765)
Tween 20
Equipment
Rocker
Stir bar and stir plate
Tripour beaker (50-mL)
METHOD
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1. Prepare BSA mix: To 25 mL of PBS, add 0.2 g of BSA. Use a 50-mL Tripour beaker, add stir bar, and add powder gradually while stirring vigorously.
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2. Prepare Tween 20 stock solution by adding 100 µL of Tween 20 to 10 mL of PBS. If using a micropipettor with disposable tips, cut off a few millimeters of the tip for a more accurate measurement. Mix on a rocker.
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3. Add 25 µL of cold water fish skin gelatin to BSA mix from Step 1.
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4. Add 0.5 mL of Tween 20 stock solution from Step 2 to the BSA/gelatin mix. Continue stirring.
- © 2012 Cold Spring Harbor Laboratory Press
