Forgot to Add Loading, Boil, Add Loading, Boil for 5 Mins, Freeze, How Much to Dilute Next Time
It is recommended to dilute the sample 2 to 5 times. The exact dilution factor depends on the sample concentration and experimental requirements. Typically, the sample should be diluted to a final protein concentration of 1-2 µg/µL to ensure the accuracy and reproducibility of electrophoresis results.
After dilution, 1X loading buffer must be added. The addition of loading buffer ensures proper migration of the sample during electrophoresis, while the bromophenol blue dye allows monitoring of the electrophoresis progress.
Following the addition of the loading buffer, the sample must be boiled for 1-2 minutes. Re-boiling ensures complete protein denaturation, preventing incomplete protein bands or abnormal migration patterns during electrophoresis.
MtoZ Biolabs, an integrated chromatography and mass spectrometry (MS) services provider.
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