Question

34. In isoelectric focusing, proteins are separated on the basis of their
a). relative content of positively charged residue only
b). relative content of negatively charged residue only
c). size
d). relative content of positively and negatively charged residue​

Answer 1

Answer:

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Answer 2

The Correct solution is Option D, proteins are separated on the basis of their relative content of positively and negatively charged residues.

• Isoelectric Focusing, or IEF, is a technique for sorting proteins in a pH gradient based on their isoelectric points.
• The pH at which proteins carry no net charge or become immobile in an electric field is known as the isoelectric point, abbreviated as pI.
• Isoelectric Focusing has a very better resolution.
• Normal electrophoretic procedures maintain a constant pH between the anode and the cathode, whereas isoelectric focusing creates a pH gradient.
• Proteins become positively charged when their pH falls below their pI, and they move towards the cathode.
• Because the charge of the protein molecule varies as it moves forward due to the pH gradient, there will be a point when the protein's net charge becomes zero, which is known as the isoelectric point.
• When a protein mixture or a single protein is exposed to an electric field at a specific pH, it ceases to move.
• Standardization of results is one of the most important aspects of isoelectric focusing. Typical protein standards were utilised for this purpose, which may have the following drawbacks:
• Solutions with a low level of stability
• Insufficient purity for use as a standard
• Inconsistency from one lot to the next
• To address these issues, we developed synthetic IEF-markers that may be utilised to detect fluorescence signals. Using fluorescent IEF-markers instead of UV detection allows for substantially higher sensitivity.