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What is the workflow of capillary electrophoresis?
Posted May 6, 2024
Answer
- Electrolyte Solution Preparation: Weigh and dissolve sodium borate in distilled water. Then adjust the pH using dilute NaOH or HCL and filter using a 0.45 μm membrane filter. Next, degas it under vacuum for 10-15 minutes.
- Capillary Preparation: Cut the 50 or 100 μm capillary and flush the capillary with 0.1 M NaOH for 30 minutes. Then rinse with distilled water for 10 minutes, 0.1 M HCl for 10 minutes and then with distilled water again for 10 minutes. Next, fill the capillary with electrolyte solution.
- Instrument Calibration: Inject standard markers and adjust the voltage and temperature for optimal separation. Input the data for the peak area and migration time for each marker.
- Sample Preparation: Mix sample with buffer and heat at 90-100°C for 5-10 minutes to denature proteins. Centrifuge at 10,000 g for 5 minutes and then inject the sample into the capillary.
- Data Analysis: Analyze electropherogram for migration time and peak area and compare them to the standard markers. Next, determine the concentration using the standard curve from the standard marker peaks.
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