Virtualab Protocols (Chang Bioscience)
1. Divide protein sample into two equal volumes, e.g.,
12.5 ml.
2. Add 2.5 ml 10X alkaline
phosphatase reaction buffer (1 M NaCl, 100 mM MgCl2,
10 mM DTT, 500 mM Tris-HCl pH7.9).
3. Add 10 ml of calf
intestinal alkaline phosphatase (CIP, 10 units/ml)
in 50% glycerol and 50 mM KCl, 1 mM MgCl2,
0.1 mM ZnCl2, 10 mM Tris-HCl pH8.2).
4. Incubate at 37 °C for 15 minutes.
5. Stop the reaction and analyze results using one of
the following methods:
-SDS-PAGE electrophoresis followed by autoradiography if the
proteins are labeled or followed by antibody detection.
-2D-gel electrophoresis followed by autoradiography if the
proteins are labeled or followed by antibody detection.
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