18 Questions
What is the purpose of removing the connections between the amino acid side chains of the antigen and the formol?
To reveal the antigenic regions in IHC application
What is the optimal temperature range for applying the solutions in the physical method of antigen unmasking?
95-100°C
Which of the following enzymes is NOT used in the chemical method of antigen unmasking?
Lipase
What is the most widely used chromogen in histochemistry?
DAB
What is the primary aim of antigen unmasking in IHC?
To reduce non-specific staining
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?
To enhance the activity of the enzyme
What is the purpose of incubating the primary antibody for 1 hour?
To allow the antibody to bind to the antigen
What is the role of DAB in immunohistochemistry?
To act as a chromogen
Why are sections treated with hydrogen peroxide?
To inactivate endogenous peroxidase
What is the purpose of using PBS solution in immunohistochemistry?
To wash and remove excess antibodies
What is the purpose of dehydrating sections through graded alcohols?
To prepare the sections for mounting
Why are slides coated with Poly-L-lysine solution?
To promote adhesion of sections
What is a major disadvantage of using avidin in the avidin-biotin method?
It can bind to negatively charged molecules
What is the primary difference between avidin and streptavidin in the avidin-biotin method?
Streptavidin has a higher binding affinity for biotin
What is the labeled antigen method used for in immunohistochemistry?
To detect antigens directly
What is the purpose of the avidin-biotin-enzyme complex in the avidin-biotin method?
To reveal the immune signal
What is the peroxidase-antiperoxidase (PAP) method used for in immunohistochemistry?
To amplify the immune signal
What is the most widely used chromogen in histochemistry?
DAB
Learn about the importance of removing chemical bonds between amino acid side chains of an antigen for Immunohistochemistry application. Understand how high temperature, alcoholic environments, or proteolytic enzymes help to reverse the structural change.
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