A student given the task of purifying a protein found that the protein had a molecular mass of 400kDa when measured by gel filtration chromatography. (This technique involves the use of a column packed with material that functions like a molecular sieve. No harsh reagents are used so it preserves the native structure of the protein. ). The student wanted to confirm the purity of the protein by SDS-PAGE. When the protein was analyzed by polyacrylamide gel electrophoresis in the presence of SDS, 3 bands with the following molecular weights were obtained: 180, 160, 60kDa. However, the student realized that he/she forgot to add 2-mercaptoethanol to the sample gel buffer when it was made up. So the student ran the gel again, this time in the presence of SDS and 2-mercaptoethanol. This time, 3 bands were observed as before, however, the molecular weights were 160, 90, and 60kDa. If you were that student what would you conclude about this set of observations?
What would be the advantage of using two independent techniques (gel filtration and SDS-PAGE) to characterize a protein?
A student given the task of purifying a protein found that the protein had a molecular mass of 400kDa when measured by gel filtration chromatography. (This technique involves the use of a column packed with material that functions like a molecular sieve. No harsh reagents are used so it preserves the native structure of the protein. ). The student wanted to confirm the purity of the protein by SDS-PAGE. When the protein was analyzed by polyacrylamide gel electrophoresis in the presence of SDS, 3 bands with the following molecular weights were obtained: 180, 160, 60kDa. However, the student realized that he/she forgot to add 2-mercaptoethanol to the sample gel buffer when it was made up. So the student ran the gel again, this time in the presence of SDS and 2-mercaptoethanol. This time, 3 bands were observed as before, however, the molecular weights were 160, 90, and 60kDa. If you were that student what would you conclude about this set of observations?
What would be the advantage of using two independent techniques (gel filtration and SDS-PAGE) to characterize a protein?
