COPY: Biochemical Techniques: SDS-PAGE

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What is the primary purpose of SDS-PAGE electrophoresis?

To separate proteins by their size (correct)

To separate proteins by their charge

To stain proteins for visualization

To separate nucleic acids by their sequence

What component is primarily responsible for providing a uniform negative charge to polypeptides in SDS-PAGE?

Polyacrylamide gel

Gel pore size

Electric field

Sodium Dodecyl Sulfate (SDS) (correct)

How does the pore size of the gel affect the SDS-PAGE results?

It determines the voltage applied

It affects the resolution of the DNA bands

It influences the mobility of proteins (correct)

It modifies the staining process

In SDS-PAGE, where do the protein bands arrange based on their molecular weight?

From cathode to anode Signup and view all the answers

What factors can influence the rate of movement of proteins during SDS-PAGE?

Gel pore size and strength of electric field Signup and view all the answers

What is a common application of SDS-PAGE besides separating proteins?

Separating DNA and RNA molecules Signup and view all the answers

What type of power supply is typically needed for running SDS-PAGE?

100-200 volts Signup and view all the answers

Which of the following is not typically part of the SDS-PAGE instrumentation?

Fluorescent lighting system Signup and view all the answers

What role does acrylamide play in SDS-PAGE electrophoresis?

It is the supporting medium that polymerizes to form polyacrylamide. Signup and view all the answers

What defines the resolving gel in SDS-PAGE electrophoresis?

It typically contains 5-10% acrylamide. Signup and view all the answers

Which statement correctly describes the stacking gel in SDS-PAGE?

It constitutes 1/3rd of the gel plate and contains 2-5% acrylamide. Signup and view all the answers

Why are different buffers used for the upper and lower reservoirs in SDS-PAGE?

To ensure different pH levels in the stacking and running gels. Signup and view all the answers

What is the primary function of Comassie Brilliant Blue R-250 in SDS-PAGE?

To detect proteins after separation by creating visible blue bands. Signup and view all the answers

What happens to the excess dye incorporated in the gel after staining?

It is removed by a specific de-staining solution. Signup and view all the answers

How does the percentage of acrylamide in the stacking gel affect protein identification?

Is inversely related to the size of the protein to be identified. Signup and view all the answers

What is the main property of the resolving gel that assists in SDS-PAGE?

It has smaller diameter pores that impact the sieving property. Signup and view all the answers

What is the primary role of SDS in SDS-PAGE electrophoresis?

To denature proteins and impart a negative charge Signup and view all the answers

Which of the following best describes reducing SDS-PAGE?

It involves the use of heat and a reducing agent to denature proteins. Signup and view all the answers

What occurs to proteins as they migrate through the SDS-PAGE gel?

Short proteins find it easier to fit through the gel pores than larger ones. Signup and view all the answers

After electrophoresis, how are the separated protein bands visualized?

Through the use of colored dyes that bind to proteins Signup and view all the answers

Which of the following is NOT an application of SDS-PAGE?

Gene cloning Signup and view all the answers

What advantage does SDS-PAGE offer over native PAGE?

It allows all proteins to migrate towards the anode. Signup and view all the answers

During the SDS-PAGE process, what is the purpose of boiling the protein mixture?

To further denature proteins and reduce disulfide linkages Signup and view all the answers

Which statement correctly describes the separation process in SDS-PAGE?

Proteins separate based on their size as they navigate the gel matrix. Signup and view all the answers