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PathCheck Applied to Measurement of Protein Solutions in the SpectraMax Plus Microplate Spectrophotometer (MaxLine Application Note #26)

rected values using the custom formula: Correct.Values*!WellPathLength ( Figure 13).


The custom formula for the Dilution column is shown in Figure 14. The term !factor refers to the sample descriptor defined in the template.


The column Final Values contains the corrected absorbance values multiplied by the dilution factor. The custom formula is Correct.Values* Dilution (Figure 15). In the Cuvette Group Table, an analogous column was created to multiply the absorbance values by the dilution factors.


Figure 16 shows the elution profiles from the gel column obtained in the plate and the cuvette. The lower plot (plate values not pathlength corrected), not surprisingly, does not strictly mirror the upper profiles because the volumes in the wells were not uniform. However, when PathCheck was applied to the plate values, the plate profile was essentially superimposable on the cuvette profile.


The above examples illustrate the utility of the PathCheck feature of the SPECTRAmax PLUS. Protein solutions can be read in a microplate and the absorbance values are identical to those obtained by reading the same solutions in a standard 1 cm cuvette. Accurate pipetting is not necessary. An additional benefit of PathCheck is that the linear range is extended above the cuvette linear range (as long as the raw absorbance values in the wells are within the linear range of the instrument).


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