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18
3. Sample Protocols
Quick Protocol:
Requires a sensitive antibody and/or higher abundant protein. All steps
performed at room temperature.
Step Cycles Time Pump Speed
Blocking 1
30 min Normal
Wash
3 3 min Normal
1°Antibody 1
60 min Normal
Wash 4
3 min Normal
2°Antibody 1
30 min Normal
Wash 4
3 min Normal
Normal Protocol:
All steps performed at room temperature.
Step Cycles Time Pump Speed
Blocking 1
60 min Normal
Wash
5 3 min Normal
1°Antibody 1
120 min Normal
Wash 5
3 min Normal
2°Antibody 1
60 min Normal
Wash 5
3 min Normal
Overnight Protocol:
For very low abundant proteins and/or antibodies with low affinity.
Step Cycles Time Pump Speed
Blocking 1
60 min Normal
Wash
5 3 min Normal
Note: alternatively, perform blocking and wash steps manually