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</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</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</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</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</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</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</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</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</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</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</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</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</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</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</p> Signup and view all the answers

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