( Theresa Redila-Flores an...)Benchmark Plus Microplate Reader Scan Well Feature

Theresa Redila-Flores and Li Tian, PhD, Bio-Rad Laboratories, 2000 Alfred Nobel Drive, Hercules, CA 94547 USA

The Benchmark Plus microplate reader is a new monochromator- based absorbance reader from Bio-Rad that offers superior convenience, accuracy, and flexibility. The Benchmark Plus is controlled by Microplate Manager software. A new feature has been designed in Microplate Manager software (version 5.1 and higher) that utilizes the monochromator design of Benchmark Plus and allows the user to prescan a well in order to determine the optimal wavelength prior to running a full plate of samples. In this note, a prescan procedure is described to show how to implement such a function into real-life applications.

In the following procedure, standard protein (bovine IgG) is added to cell culture medium without serum and the Scan Well function in Benchmark Plus is used to determine the best wavelength for protein assay with cell culture medium. Bio-Rads protein assay kit (catalog # 500-0001) is used to check for measurement inconsistencies in culture medium.

Procedure
1. Serially dilute (1:2) standard bovine IgG protein ranging from 62.5 g/ml to 1 mg/ml in F-12 Hams medium without serum.

2. Aliquot 10 l (in triplicate) of each diluted standard protein into a 96-well clear bottom black microplate; i.e., A1, B1, and C1 for 1 mg/ml protein sample. Aliquot 10 l (in triplicate) of F-12 Hams medium into 3 wells (A7, B7, C7) to be used as blanks.

3. Add 200 l of diluted dye reagent (from Bio-Rad protein assay kit) to each well. Mix thoroughly using a microplate mixer for about 5 sec. Make sure that no bubbles are present. Bubbles may cause incons
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