Labeling of Antibodies Flashcards
Different Methods of Labeling Antibodies
- Enzymatic Method
- Fluorochrome Labeling
- Use of Colloidal Metals
- Radio label/Radioactive Isotopes
Enzymes that are being used for Enzymatic labeling of antibodies
- Horse Radish Peroxidase
- Alkaline Phosphatase
Most commonly used enzyme to label antibodies
Horse Radish Peroxidase
Alternative enzyme for HRP
Alkaline Phosphatase
If enzymes are used to label antibodies it must have ____
Chromogen
Chromogen are used as
Color developer (antibody-antigen reaction)
Chromogen that produces brown color
3,3’ DAB
3,3’ Diaminobenzidine
Chromogen that produces Brick red color
3-amino-9-ethylcarbazole (AEC)
If chromogens are used then it needs ____
Substrate
Substrate for HRP
Hydrogen peroxide
Substrate for ALP
Phosphate
Microscope used in Enzymatic method
Light Microscope
Advantage of Enzymatic Method to label antibodies
- long storage life
- Hematoxylin as counterstain
- Uses light microscope
- Fluorescence Microscope is not used
Fluorochrome labeling uses
FIC (Fluorescein Isothiocyanate)
Fluorescence Microscope
Used of colloidal metals uses
Gold (to label)