Antigen Preparation for Immunohistochemistry
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Questions and Answers

What is the purpose of removing the connections between the amino acid side chains of the antigen and the formol?

  • To enhance the staining process
  • To increase the binding capacity of the antigen
  • To reveal the antigenic regions in IHC application (correct)
  • To reduce the temperature of the solution

    • What is the optimal temperature range for applying the solutions in the physical method of antigen unmasking?

  • 100-105°C
  • 90-95°C
  • 95-100°C (correct)
  • 80-90°C

    • Which of the following enzymes is NOT used in the chemical method of antigen unmasking?

  • Pepsin
  • Trypsin
  • Proteinase K
  • Lipase (correct)

    • What is the most widely used chromogen in histochemistry?

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

    What is the primary aim of antigen unmasking in IHC?

    <p>To reduce non-specific staining</p> Signup and view all the answers

    What is the purpose of incubating the sample at 37°C for 10-20 minutes in a humid box during the chemical method of antigen unmasking?

    <p>To enhance the activity of the enzyme</p> Signup and view all the answers

    What is the purpose of incubating the primary antibody for 1 hour?

    <p>To allow the antibody to bind to the antigen</p> Signup and view all the answers

    What is the role of DAB in immunohistochemistry?

    <p>To act as a chromogen</p> Signup and view all the answers

    Why are sections treated with hydrogen peroxide?

    <p>To inactivate endogenous peroxidase</p> Signup and view all the answers

    What is the purpose of using PBS solution in immunohistochemistry?

    <p>To wash and remove excess antibodies</p> Signup and view all the answers

    What is the purpose of dehydrating sections through graded alcohols?

    <p>To prepare the sections for mounting</p> Signup and view all the answers

    Why are slides coated with Poly-L-lysine solution?

    <p>To promote adhesion of sections</p> Signup and view all the answers

    What is a major disadvantage of using avidin in the avidin-biotin method?

    <p>It can bind to negatively charged molecules</p> Signup and view all the answers

    What is the primary difference between avidin and streptavidin in the avidin-biotin method?

    <p>Streptavidin has a higher binding affinity for biotin</p> Signup and view all the answers

    What is the labeled antigen method used for in immunohistochemistry?

    <p>To detect antigens directly</p> Signup and view all the answers

    What is the purpose of the avidin-biotin-enzyme complex in the avidin-biotin method?

    <p>To reveal the immune signal</p> Signup and view all the answers

    What is the peroxidase-antiperoxidase (PAP) method used for in immunohistochemistry?

    <p>To amplify the immune signal</p> Signup and view all the answers

    What is the most widely used chromogen in histochemistry?

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

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