Buffers and pH

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The page below is a sample from the LabCE course Electrophoresis. Access the complete course and earn ASCLS P.A.C.E.-approved continuing education credits by subscribing online.

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Buffers and pH

The isoelectric point of most proteins is between pH 4.0 and 7.5. In pH 8-9, proteins will take on a negative charge and migrate to the anode. Most protein electrophoresis is performed at pH 8.6.
Buffers most commonly used are barbital or tris-boric acid-EDTA buffers. They fix the pH at 8.6, leading to sharper bands and good separations.