Microplate Procedure for Detection of Horseradish Peroxidase

1.1     96-well microplate, black or white opaque

1.2    HRP at 0.4 – 25 ng/ml

1.3    Buffer for HRP dilution

1.1     Per insert instruction, mix equal parts of chemiluminescent substrate solution A and solution B.  Store in an amber bottle.

1.2     Dispense 10 µl of HRP solution into the microwell.

1.3     Add 100ul of mixed chemiluminescent substrate solution.

1.4     Place the microplate on a luminometer and measure photocount (relative light units) between 1- 5 minutes.

ELISA Microplate Procedure

1.1     96-well microplate, black or white opaque

1.2     Diluted capture antibody

1.3     Wash buffer

1.4     Block buffer

1.5     Analyte

1.6     HRP-Antibody conjugate

2.1    Follow standard ELISA procedures.  Coat, incubate, block and wash microplate.

2.2    Dispense analyte.  Incubate and wash microplate.

2.3    Add HRP conjugate.  Incubate and wash microplate.

2.4    Per insert instruction, mix equal parts of chemiluminescent substrate solution A and solution B. Store in an amber bottle.

2.5    Add mixed chemiluminescent substrate solution to each well.

2.6    Mix liquid in wells if required.

2.7    Place the microplate on a luminometer and measure photocount (relative light units) between 1-5 minutes

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