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Questions and Answers
What determines the migration speed of proteins in serum protein electrophoresis?
What determines the migration speed of proteins in serum protein electrophoresis?
Which medium condition gives proteins a negative charge in serum protein electrophoresis?
Which medium condition gives proteins a negative charge in serum protein electrophoresis?
Which buffer is used in serum protein electrophoresis to ensure proteins are negatively charged?
Which buffer is used in serum protein electrophoresis to ensure proteins are negatively charged?
After separation, which dyes are commonly used to stain the proteins on the cellulose acetate strip?
After separation, which dyes are commonly used to stain the proteins on the cellulose acetate strip?
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What is the purpose of using 5% acetic acid in serum protein electrophoresis?
What is the purpose of using 5% acetic acid in serum protein electrophoresis?
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How many bands are typically observed on a cellulose acetate strip after serum protein electrophoresis?
How many bands are typically observed on a cellulose acetate strip after serum protein electrophoresis?
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Study Notes
Serum Protein Electrophoresis (SPE)
- Proteins are amphoteric molecules, meaning they can be uncharged, negatively charged, or positively charged depending on the pH of the surrounding media.
Protein Charge and pH
- In alkaline media (high pH), proteins are negatively charged (anions).
- In acidic media (low pH), proteins are positively charged (cations).
- In neutral media (pH 7), proteins are neutral.
Principle of SPE
- SPE separates and semi-quantifies serum proteins using an electrical field in alkaline media (Barbital buffer, pH 8.6).
- Negatively charged protein molecules migrate towards the positive electrode (anode) at varying speeds based on:
- Number of negative charges they carry (more negative charge, faster migration).
- Size and shape of the molecule.
SPE Process
- Serum protein mixture is separated into distinct bands, typically 5 bands on a cellulose acetate strip.
- The strip is then stained with dyes like Ponceau S or amido black.
- The background is cleared with 5% acetic acid.
- The strip is dried and interpreted by eye and densitometer.
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Description
Learn about the basics of Serum Protein Electrophoresis (SPE), including protein charge and its relation to pH levels. Understand how SPE separates and semi-quantifies serum proteins.
