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SDS-PAGE Gel Electrophoresis

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Questions and Answers

What is the primary purpose of treating samples with protease inhibitor during protein extraction?

To increase protein yield
To enhance protein solubility
To prevent protein denaturation (correct)
To remove impurities from the sample

What is the main difference between Southern blotting and Western blotting?

Type of antibody used
Molecular weight of the target molecule
Type of gel used for separation
Target molecule being detected (DNA vs. protein) (correct)

What is the purpose of the blocking step in Western blotting?

To enhance antibody binding
To increase protein transfer efficiency
To prevent non-specific binding (correct)
To reduce background noise

What is the primary function of the secondary antibody in Western blotting?

To enhance signal detection (D) Signup and view all the answers

What is the purpose of SDS-PAGE in Western blotting?

To separate proteins based on molecular weight (D) Signup and view all the answers

What is the mechanism of protein transfer from the gel to the membrane in Western blotting?

Electrotransference (B) Signup and view all the answers

What is the primary function of glycerol in the loading buffer during protein extraction?

To help sink the sample in the well (C) Signup and view all the answers

What is the driving force behind the transfer of proteins from the gel to the nitrocellulose membrane during electro-blotting?

Electric current (A) Signup and view all the answers

What is the primary purpose of blocking in western blotting?

To prevent non-specific binding of antibodies to the nitrocellulose membrane (D) Signup and view all the answers

What is the primary factor that determines the separation of proteins in SDS-PAGE?

Molecular weight and electric charge (A) Signup and view all the answers

What is the advantage of using electro-blotting over traditional blotting?

Reduced time required for protein transfer (A) Signup and view all the answers

What is the role of bromothymol blue in the sample preparation for SDS-PAGE?

To monitor the movement of proteins during electrophoresis (C) Signup and view all the answers

What is the primary function of the secondary antibody in a Western blotting procedure?

To recognize and bind to the primary antibody (A) Signup and view all the answers

Which of the following enzymes is NOT typically used as a label for the secondary antibody in Western blotting?

Lactate dehydrogenase (D) Signup and view all the answers

What is the purpose of incubating the reaction mixture with a specific substrate in Western blotting?

To visualize the enzyme action (B) Signup and view all the answers

What is a primary application of Western blotting in disease diagnosis?

Detection of antibodies against viruses or bacteria in serum (C) Signup and view all the answers

What is the confirmatory test for HIV that utilizes Western blotting?

Detection of anti-HIV antibodies in patient's serum (A) Signup and view all the answers

What is the type of protein that can be detected using Western blotting?

All types of proteins (A) Signup and view all the answers

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