5 Questions
What is the pH of the barbital buffer used in the electrophoresis chamber for serum protein electrophoresis?
8.6
Which dye is commonly used for staining the serum protein bands in SPE?
Ponceau S
After the electrophoresis step, what is the next step in the serum protein electrophoresis procedure?
Applying the strip to stain
During the destaining step of serum protein electrophoresis, the strip is sequentially applied in containers with what concentration of acetic acid?
5%
What method is used for semiquantitative estimation of each separated band in serum protein electrophoresis?
Visual comparison with a normal serum band
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.
Steps involved in Serum Protein Electrophoresis (SPE), a laboratory technique used to separate and identify serum proteins. Learn about the application, electrophoresis, and separation of proteins into distinct bands.
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