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The corrected mP value is not calculated correctly for Fluorescent Polarisation

Document ID: DQ076
Relevant to following products: WorkOut 2.0
Document description: How To: Set-up a Fluorescent Polarisation with corrected mP

To set-up Fluorescent Polarisation assay use the Generic Wizard and set-up the VICTOR measurements as normal. Then take measurements as required.

If you have a blank group on the microplate layout and use WorkOut’s Generic wizard, it will setup a blank correction transform for each raw matrix. Thus with FP measurements since, you get 3 raw matrices (S,P and mP), the wizard adds 3 blank corrected matrices. However, with FP the corrected mP should be calculated using the equation:

mP(Corr) = 1000 * [S(Corr) - P(Corr)] / [S(Corr) + P(Corr)]

Therefore to use this equation instead you must remove the 3rd blank correction transform added by the Generic wizard and add the correct equation. To do this, select Edit Transforms, first remove the %CV transform if it is present, select the 3rd Blank Correction transform and click Remove. Now press Create a new transform and select Matrix Expression. Under the Expression tab enter the Expression:

1000 * (w.4.1 - w.5.1) / (w.4.1 + w.5.1)

This corresponds to the equation above. Also, under the Output tab enter a matrix name such as mP(Corr).

Related articles:
DQ077: How can I see the mP during Fluorescent Polarisation measurements?
How can I see the mP during Fluorescent Polarisation measurements?

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