What is indirect immunocytochemistry?
Answers
Answer:
Direct IF uses a single antibody directed against the target of interest. The primary antibody is directly conjugated to a fluorophore. Indirect IF uses two antibodies. The primary antibody is unconjugated and a fluorophore-conjugated secondary antibody directed against the primary antibody is used for detection.
Answer:
Indirect immunofluorescence, or secondary immunofluorescence, is a technique used in laboratories to detect circulating autoantibodies in patient serum. It is used to diagnose autoimmune blistering diseases.
Unlabelled primary antibodies (used as a marker for cancer, diabetes and Alzheimer disease) from the patient serum bind to the target molecule in pre-prepared tissue samples.
Secondary antibodies (used as a marker for HIV), conjugated with a fluorescent dye such as fluorescein isothiocyanate (FITC), bind to the primary antibody.
When exposed to light, the fluorescent dye is excited and emits a wavelength that can be seen with a fluorescent microscope.
Explanation: