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Chapter 32 : Electrophoresis of Amylase Samples
This chapter talks about the separation of the proteins in amylase samples, such as saliva, by electrophoresis and the identification of the amylase bands. Electrophoresis of DNA is a little different from electrophoresis of proteins. The differences are due to the fact that proteins are quite different from DNA. DNA has a uniform backbone that is negatively charged at the pH of electrophoresis buffers. Therefore, the major difference between DNA fragments is usually their size. During standard electrophoresis, they all migrate toward the positive pole at rates that depend on their sizes. The net effect is that the proteins all migrate toward the positive pole during electrophoresis (as do DNA molecules). After electrophoresis, the gel is cut in half so that two halves are obtained, each with a complete sample set. After electrophoresis, half the gel is stained for total protein, and the other half is assayed for amylase activity. The gel is removed from the electrophoresis chamber, and cut in half vertically with a razor blade, ruler edge, or scissor blade. Many of the complex samples, such as bean extract or human saliva, show multiple bands. To determine which of these bands is amylase, the unstained half of the gel for areas of visible clearing is carefully examined.
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