Yu:Western

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Western Blotting

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SDS PAGE

1.Cast SDS-PAGE gel and assemble the gel apparatus.
2. Add 10μL of desired sample and 5μL 3x Gel loading buffer to an eppendorf tube and boil for 5 minutes. Do this in separate tubes for each sample to be analyzed. After boiling, samples may be kept at room temperature until loaded
3. Load 12μL to 15μL of sample into each lane. Load protein prestain marker, as well as BenchMark, if desired. Load any empty lanes with 1x Gel loading buffer
4. Run gel until the loading buffer is just running off the bottom of the gel

Transfer

Immunoblotting

Developing