SDS-polyacrylamide gel electrophoresis (PAGE) procedure
1. Install the glass plate as shown
2. Prepare the separation gel (10%) according to the table, mix the separation gel and immediately inject into the gap of the glass plate. Carefully cover the upper layer with a layer of ethanol, and place the rubber plate vertically at room temperature. Go to the top of the ethanol and blot dry with filter paper.
3. Preparation of concentrated gel (4%): The concentrated gel was mixed and directly poured onto the polymerized separating gel, and the comb was immediately inserted, and the gel was vertically placed at room temperature for polymerization.
4. Sample treatment: When the concentrated gel is polymerized, the sample is mixed with an equal volume of sample buffer and heated at 100 ° C for 1-2 min.
5. After the polymerization of the concentrated gel is complete, carefully remove the comb, then fix the rubber plate on the electrophoresis device, and add the electrophoresis buffer to each of the upper and lower tanks.
6. Loading: Load with a micro-injector and add 20 μl to each sample.
7. Electrophoresis: Electrophoresis was carried out at a constant pressure of 100-150 V until the bromophenol blue reached the bottom of the gel and the power was turned off.
8. Staining: Remove the glass plate from the electrophoresis device, carefully remove the gel from the slide glass plate, and stain it in the staining solution for more than 2 hours.
9. Decolorization: Remove the gel and put it into the decolorizing solution to decolorize until the background is colorless.
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