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?

<p>To enhance signal detection (D)</p> Signup and view all the answers

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

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

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

<p>Electrotransference (B)</p> Signup and view all the answers

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

<p>To help sink the sample in the well (C)</p> 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?

<p>Electric current (A)</p> Signup and view all the answers

What is the primary purpose of blocking in western blotting?

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

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

<p>Molecular weight and electric charge (A)</p> Signup and view all the answers

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

<p>Reduced time required for protein transfer (A)</p> Signup and view all the answers

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

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

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

<p>To recognize and bind to the primary antibody (A)</p> 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?

<p>Lactate dehydrogenase (D)</p> Signup and view all the answers

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

<p>To visualize the enzyme action (B)</p> Signup and view all the answers

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

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

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

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

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

<p>All types of proteins (A)</p> Signup and view all the answers

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