Liquid Phase Isoelectric Focusing: An Electrophoresis Technique to Separate Bioactive Molecules from Crude Gymnema sylvestre Extract
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Add 0.6 grams of Gymnema sylvestre plant extract to 60 milliliters of distilled water. Dissolve the plant extract by mixing in a roller tube for five minutes.
To remove the insoluble particles, centrifuge the solution at 10,000 x g for five minutes. Transfer the supernatant to a 150-milliliter glass beaker, and add ampholyte to create a 1% solution by volume, approximately 0.6 milliliters.
Using a 50-milliliter syringe, with a 1.5-inch, 19-gauge blunt-end needle, load the solution into the IEF cell through the sample collection ports. Then, remove the cell from the stand, and tap the electrode chamber to dislodge and remove any bubbles.
Connect the unit to the cooling water. With the power supplied at constant 15 watts, begin fractionation. Run the IEF unit until the voltage reaches a constant value, approximately three hours.
After pressing the Harvest ON button, align the fraction collector pins with the collection ports. Push the pins through the sealing tape to begin collecting the Gymnema sylvestre fractions.
