An Introduction to SDS Page Flashcards
What is the name of this lab?
An Introduction to SDS Page
What is electrophoresis?
To carry with electricity
General Principles of Protein Electrophoresis and __________ is the migration of charged molecules in an electric field toward the electrode with the _____ charge.
SDS-PAGE Electrophoresis; the opposite
What is the acronym SDS-PAGE stand for?
Sodium dodecyl sulfate-polyacrylamide gel electrophoresis
SDS-PAGE is a form of electrophoresis that treats samples with ____ to ____ proteins.
SDS; denature
What value is there on the SDS-PAGE technique?
It helps find the:
- # proteins in a sample;
- molecular weights of proteins;
- differences in proteins from different sources;
- purity of protein of interest;
- amount of protein present
Proteins are usually separated using agarose gels. True or False?
False. Proteins are separated using polyacrylamide gels. (Agarose gels are used to separate DNA.)
Proteins are usually separated using polyacrylamide gels. True or False?
True
Why are proteins separated by polyacrylamide gels?
Most proteins are much smaller than DNA fragments AND polyacrylamide gels have pore sizes similar to the sizes of proteins.
The gel matrix formed by polyacrylamide is much ______ than agarose and able to resolve much _____ molecules.
tighter; smaller
Polyacrylamide gels are often used to separate very _____ molecules (_____ bp) for DNA sequencing or PCR analysis, while agarose is sometimes used to separate very ____ molecules.
small; <500 bp; large
PAGE uses how many phases of polyacrylamide?
2
The two phases of polyacrylamide used are the upper stacking gel typically of __% acrylamide and a lower resolving gel of a ____ percentage of acrylamide.
4; higher
This is called a _______ system and results in all of the proteins in a sample ______, or ______, at the same time.
discontinuous; separating; resolving
Since sample volumes can vary from lane to lane, forming vertically narrow or broad bands in the wells, all of the proteins in a sample enter the gel simultaneously. True or False.
False, The proteins do NOT enter the gel simultaneously.
The ___ percentage of the stacking gel allows the proteins to migrate _____ and be _____ at the edge of the denser resolving gel regardless of their sizes.
low; rapidly; compressed
The samples of _________ are thus concentrated ____ thin bands in each lane before they move into the denser resolving gel and begin to be separated according to their ________.
mixed proteins; uniformly; molecular weights
To establish the ion fronts, the SDS-PAGE running buffer is made with _____ and ____ (ph ___)….
Tris; glycine; 8.3
The Ready Gel polyacrylamide gel is made with _____ (pH ____).
Tris-HCl; 8.8.
Since ____ ions migrate more rapidly than ____ ions in an electric field, and proteins have intermediate mobility; the proteins become _____ in a narrow band between ion fronts when electrophoresis is begun.
chloride; glycine; trapped
Polyacrylamide does not require a chemical reaction to cause polymerization of two acrylamide monomers. True or False?
False. They do require a chemical reaction.
Polyacrylamide requires a chemical reaction to cause polymerization of two acrylamide monomers. True or False?
True
What must be added to a solution containing the desired concentrations of acylamide and bis-acrylamide monomers in a Tris buffer to cast a polyacrylamide gel?
- a reaction inhibitor;
- ammonium persulfate (APS);
- catalyst
- tetramethylethylenediamine (TEMED)
To cast a gel with a resolving and stacking gel of different polyacrylamide concentrations, a ____________ gel is poured first and the ____ concentration stacking ___ is poured on top of it.
high-concentration resolving; low; gel
