VMC 221: Veterinary Immunology and Serology (1+1)

IMMUNOFLUORESCENCE ASSAYS

• Fluorescent dyes are commonly used as labels in primary binding tests. The most important and commonly used dye is FITC (fluorescein isothiocyanate). Rhodamine isothiocyanate is also used.
• FITC is a yellow compound that can be bound to antibodies without affecting their reactivity.
• When irradiated with UV light, FITC re- emits visible green/or yellow green light at 525 m m.
• Rhodamine emits deep red fluorescence (546 m m) and phycoerythrin emits red fluorescence.
• Three types of immunofluorescent assays are there,
  • Direct
  • Indirect
  • Sandwich