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# Antibody is adsorbed onto solid phase
# Antibody is adsorbed onto solid phase
# Wash
# Wash
# Sample is added- specific antigen binds to antibody
# Sample serum is added- specific antigen binds to antibody
# Wash
# Wash
# Enzyme-labeled specific antibody is added- attaches to bound antigen
# Enzyme-labeled specific antibody is added- attaches to bound antigen
# Wash
# Wash
# Enzyme substrate is added
# Enzyme substrate is added
''visualised product = amount of antigen in the sample''
''visualised product = amount of antigen in the serum''
'''Antibody Class Capture Assay''' (for antibody detection)
'''Antibody Class Capture Assay''' (for antibody detection)
# Class-specific antiglobulin is adsorbed onto solid phase
# Class-specific antiglobulin is adsorbed onto solid phase
# Wash
# Wash
# Sample is added- class-specific antibody in the sample binds to antiglobulin
# Sample serum is added- class-specific antibody in the serum binds to antiglobulin
# Wash
# Wash
# Antigen is added- attaches to specific antibody
# Antigen is added- attaches to specific antibody
# Wash
# Wash
# Enzyme substrate is added
# Enzyme substrate is added
''visualised product = amount of specific antibody in sample''
''visualised product = amount of specific antibody in serum''
'''Indirect Method''' (for antibody detection)
'''Indirect Method''' (for antibody detection)
# Antigen is adsorbed onto solid phase
# Antigen is adsorbed onto solid phase
# Wash
# Wash
# Sample is added- any specific antibody present in sample binds to antigen
# Serum is added- any specific antibody present in sample binds to antigen
# Wash
# Wash
# Enzyme-labeled antiglobulin is added- attaches to antibody
# Enzyme-labeled antiglobulin is added- attaches to antibody
# Wash
# Wash
# Enzyme substrate is added
# Enzyme substrate is added
''visualised product = amount of antibody in the sample''
''visualised product = amount of antibody in the serum''
===Competitive ELISA===
===Competitive ELISA===
# Antigen is adsorbed onto solid phase
# Antigen is adsorbed onto solid phase
# Wash
# Wash
# Enzyme-labeled antibody (pre-titrated- optimal colour development) and sample are added
# Enzyme-labeled antibody (pre-titrated- optimal colour development) and serum are added
# Wash
# Wash
# Enzyme substrate is added
# Enzyme substrate is added
# Antigen is adsorbed onto solid phase
# Antigen is adsorbed onto solid phase
# Wash
# Wash
# Antigen in sample is incubated with enzyme-labelled antibody (again pre-titrated): this is directed against the antigen on the solid phase
# Antigen in serum is incubated with enzyme-labelled antibody (again pre-titrated): this is directed against the antigen on the solid phase
# Wash
# Wash
# Enzyme substrate is added
# Enzyme substrate is added
''visualised product = amount of enzyme-labelled antigen''
''visualised product = amount of enzyme-labelled antigen''