1. Although you are making 12% SDS – PAGE gels, other percentages can be made. a....
1. Although you are making 12% SDS – PAGE gels, other percentages can be made.
a. When would you want to use a 10% gel?
b. When would you want to use a 20% gel?
2. Why must you use both acrylamide and bis – acrylamide to make
your gel?
3. There are three ratios of acrylamide:bis – acrylamide that are
used when making these
types of gels.
a. What is the primary application for a gel made up of 19:1
acrylamide:bis –acrylamide?
b. What is the primary application for a gel made up of 29:1
acrylamide:bis –acrylamide?
c. What is the primary application for a gel made up of 37.5:1
acrylamide:bis –acrylamide?
Homework Answers
Answer: 1
a) The combination of SDS-PAGE with a protein stain is used in biochemistry for the quick and exact separation of proteins. It is also used for and subsequent analysis of proteins.
b) This SDS-PAGE is mainly used in mass spectrometry of proteins for the sample preparation in-gel digestion.
Answer: 2 :
Acrylamide is used as precursor for the preparation of electrophoretic gels which are mainly used to separate proteins by size. Acrylamide mixed with bisacrylamide to form a cross-linked polymer network by the addition of polymerizing agent, ammonium persulfate (APS).
The reaction can be catalyzed by TEMED (N,N,N,N'-tetramethylenediamine) that promotes the production of free radicals by APS. Here polyacrylamide gel acts as as a support medium and sodium dodecyl sulfate (SDS) is used to denature the proteins.
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1. Although you are making 12% SDS – PAGE gels, other
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