1. Greatest Amplification:
Legend:
Antigen
Biotin
Antigen
Avidin
Step 1
Primary Antibody
(with light and heavy chains)
(use BSA)
Enzyme
Step 2
Biotinylated Secondary Antibody
(use BSA)
Avidin (Neutravidin, Extravidin, Streptavidin) with
Step 3
(use TTBS)
Enzyme (HRP - horseradish peroxidase )
Step 4
ECL
Light
Lesser Amplification:
Antigen
Antigen
Primary Antibody
(with light and heavy chains)
(use BSA)
Primary Antibody
(with biotin)
(use BSA)
Step 1
Step 1
Avidin with
Secondary Antibody
with HRP
**do not use BSA (TTBS)
Step 2
(use TTBS)
Step 2
HRP
Step 3
Step 3
ECL
Light
ECL
Light