18 Questions
What is the purpose of immunofluorescence?
To detect antigens in cellular contexts using antibodies.
What is the difference between indirect and direct immunofluorescence?
In indirect immunofluorescence, a secondary fluorescent antibody binds to the primary antibody, while in direct immunofluorescence, only one marked primary antibody is incubated without a second antibody.
List the protocol types involved in immunofluorescence.
Tissue or cell preparation, tissue or cell fixation, serum blocking, primary antibody incubation, marked or fluorophore second antibody incubation, staining, result judgment, imaging.
What is the mechanism of indirect immunofluorescence?
Primary antibodies directly bind to antigen, then a secondary fluorescent antibody binds to the primary antibody.
What is the main difference between direct and indirect immunofluorescence?
The main difference is in the number of antibodies used: direct uses one antibody directed against the target, while indirect uses two antibodies.
What are the advantages of the direct immunofluorescence method over the indirect method?
The direct method is advantageous in terms of shorter duration, reduced cost, and simplicity.
What are the advantages of the indirect immunofluorescence method over the direct method?
The indirect method offers advantages in terms of flexibility, sensitivity, and species cross reactivity.
How does the indirect immunofluorescence method achieve signal amplification?
The signal amplification is achieved by several secondary antibodies binding to the primary antibody, resulting in an amplified signal.
What is the main advantage of using pre-conjugated primary antibodies in immunofluorescence?
The advantage is in minimizing non-specific binding, as the fluorophore is already conjugated to the primary antibody.
How does the direct immunofluorescence method minimize species cross reactivity?
Direct immunofluorescence minimizes species cross reactivity by using primary antibodies with the fluorophore already conjugated.
What is the process of immunoelectrophoresis?
Immunoelectrophoresis is a technique that involves precipitating proteins in agar under the influence of an electric field, separating the antigen mixture into component elements, and reacting antigens with specific antiserum to form precipitin lines.
Who coined the term 'Immunoelectrophoresis' and in what year?
The term 'Immunoelectrophoresis' was created by Grabar and Williams in 1953.
What is the principle of immunoelectrophoresis?
The principle involves separating the antigen mixture into single antigen components based on their size and charge, followed by the reaction of antigens with specific antiserum to form precipitin lines.
What is the advantage of using indirect immunofluorescence method over the direct method?
Signal amplification
How does the direct immunofluorescence method minimize species cross reactivity?
Minimized by the fluorophore already conjugated to the primary antibody
What is the main difference between direct and indirect immunofluorescence?
Number of antibodies used
What is the process of immunoelectrophoresis?
A technique to characterize antibodies
Who coined the term 'Immunoelectrophoresis' and in what year?
Grabar and Williams in 1953
Test your knowledge about immuno-fluorescence and immuno-electrophoresis, including the assay used primarily on biological samples and the procedure to detect antigens in cellular contexts using antibodies. Explore the types, principles, and protocols of indirect immuno-fluorescence.
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