Hoatlin:Western Protocol
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BASIC WESTERN BLOT PROTOCOL
• Block membrane 1 hour in blocking buffer (5% carnation non-fat dry milk dissolved in water.)
• Incubate membrane for 1 hour in blocking buffer containing primary antibody.
• Wash the membrane 2X5 min, 2X10 min in 0.05% Tween 20 in TBS.
• Incubate membrane for 30min in blocking buffer containing secondary antibody with desired (HRP, AP, etc.) conjugate.
• Wash the membrane 2X5 min, 2X10 min in 0.05% Tween 20 in TBS.
• Use ECL according to manufacturer to develop.
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