Critical Considerations and Actual Trends
Some major drawbacks of the common protocols for the in-gel digestion are the extended time need and the multiple processing steps making the method error-prone in respect to contaminations (especially keratin). These disadvantages were largely removed by the development of optimised protocols and specialised reaction tubes.
More severe than the difficulties with handling are losses of material while processing the samples. The mass spectrometric protein analysis is often performed at the limit of detection, so even small losses can decide about success or failure of the whole analysis. These losses are due to washout during different processing steps, adsorption to the surface of reaction tubes and pipette tips, incomplete extraction of peptides from the gel and/or bad ionisation of single peptides in the mass spectrometer. In dependence of the different physicochemical properties of the peptides the losses can vary between 15 and 50%. Due to this heterogeneity of the peptides up to now a universally valid solution for this major drawback of the method has not been found.
Read more about this topic: In-gel Digestion
Famous quotes containing the words critical, actual and/or trends:
“The principle goal of education in the schools should be creating men and women who are capable of doing new things, not simply repeating what other generations have done; men and women who are creative, inventive and discoverers, who can be critical and verify, and not accept, everything they are offered.”
—Jean Piaget (1896–1980)
“We can never safely exceed the actual facts in our narratives. Of pure invention, such as some suppose, there is no instance. To write a true work of fiction even is only to take leisure and liberty to describe some things more exactly as they are.”
—Henry David Thoreau (1817–1862)
“A point has been reached where the peoples of the Americas must take cognizance of growing ill-will, of marked trends toward aggression, of increasing armaments, of shortening tempers—a situation which has in it many of the elements that lead to the tragedy of general war.... Peace is threatened by those who seek selfish power.”
—Franklin D. Roosevelt (1882–1945)