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

t fit properly.
Disposable UV Plate, Corning Costar Corp. (catalog # 3635)
UV-MAX, (Polyfiltronics, Inc., catalog # VO32SCXP8)

3. Pipettor and tips or transfer pipets suitable for use with microplates.

4. Samples (100-300 L each)

5. Quartz cuvette


Set up to make pre-read measurements
Step 1 Launch SOFTmax PRO, then open a Plate section or, if necessary, create a new Plate section.

Step 2 Set up the Instrument Settings dialog box as shown in Figure 1. In the Instrument Settings dialog box, select to perform an endpoint read at 280 nm. Select PathCheck, (Pre-read plate will automatically be checked) and select Cuvette Reference. (Alternatively, you can use the factory-installed Water Constant as a shortcut, though it is potentially less accurate.)


Step 3 Use the Template Editor to create a template showing where standards, blanks, and unknowns will be located on the microplate.

Step 4 Set up the Reduction dialog box as shown in Figure 2. Set the wavelength combination to L1 and the data mode to Absorbance. Make sure that Use pre-read plate and Apply Pathcheck are selected.


METHOD
Making pre-read measurements
The purpose of the pre-read is to subtract the background absorbance of the plate. In contrast, blanks (plate, sample or group) are designated in the template and are subtracted after pathlength correction. In some cases, you may choose to use stored pre-read values (See below).
Step 1 Select a clean microplate and pipet distilled
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