Serum Protein Electrophoresis (SPE) Steps

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Questions and Answers

What is the pH of the barbital buffer used in the electrophoresis chamber for serum protein electrophoresis?

  • 9.0
  • 6.8
  • 8.6 (correct)
  • 7.4

Which dye is commonly used for staining the serum protein bands in SPE?

  • Safranin
  • Ponceau S (correct)
  • Coomassie Brilliant Blue
  • Methylene Blue

After the electrophoresis step, what is the next step in the serum protein electrophoresis procedure?

  • Scanning the strip with a densitometer
  • Applying the strip to stain (correct)
  • Applying the strip to a barbital buffer
  • Drying the strip

During the destaining step of serum protein electrophoresis, the strip is sequentially applied in containers with what concentration of acetic acid?

<p>5% (D)</p> Signup and view all the answers

What method is used for semiquantitative estimation of each separated band in serum protein electrophoresis?

<p>Visual comparison with a normal serum band (B)</p> Signup and view all the answers

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Study Notes

Steps of Serum Protein Electrophoresis (SPE)

  • Patient and normal control serum are applied to a supporting media, such as paper, agarose gel, or cellulose acetate, with cellulose acetate being the most commonly used.
  • The strip containing serum is then placed in an electrophoresis chamber filled with a barbital buffer of pH 8.6.
  • An electrical field is applied to the chamber for a constant time and voltage to separate serum proteins into distinct bands.
  • The strip is removed from the chamber and stained with a dye, typically Ponceau S or amido black.
  • The strip is then destained by sequential treatment in 5% acetic acid containers until the background is cleared.
  • The strip is dried using a special filter paper or in an oven.
  • The separated bands (typically 5 bands) are semiquantitatively estimated by densitometer scanning and visual comparison with a normal serum band.

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