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)
Gold in Colloidal method of labelling produces what color
PINK color product
Colloidal Method of labeling uses what microscope
Light Microscope
Other than Antibody, what substance can be used to identify antigen
Lectins
It can bind carbohydrates and tissue
Lectins
Source of Lectins
Plants or Animals
Incubation Requirement for IHC
30-60mins @Room temperature
