Western Blot (AP)
主要试剂
1. Membrane Blocking buffer:5% Milk 0.05% of Tween 20 PBS
2. antibody dilution buffer: PBS 0.05% of Tween20 1.0% Milk(or BSA)
3. washing buffer: PBS 0.05% of Tween20
实验步骤
Western Blot (AP)
1. Membrane Block
5% Milk 0.05% of Tween 20, Sharking for 2hr
Wash the membrane 3X with PBS(0.05% of Tween 20), 10min each
Primary antibody
Add primary antibody (antibody mix with PBS 0.05% of Tween20 1.0% Milk(or BSA) with proper dilution), 4◦C, O/N.
Wash the membrane 3X with PBS(0.05% of Tween 20), 10min each
Secondary antibody
Add second antibody (antibody mix with PBS 0.05% of Tween 20, for AP) RT,
1 hr.
Wash the membrane 3X with PBS(0.05% of Tween 20), 10min each
7. Exposure the membrane in AP buffer.
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综述
