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Benchmark Plus Microplate Reader Scan Well Feature

istencies in absorbance reading.

4. Incubate 5 min at room temperature.

5. Place microplate into Benchmark Plus microplate reader.

6. Open Microplate Manager. Under File, select Endpoint protocol.

7. Select 96-well format. Under Advanced Options, open Scan Well.

8. Scan well A1 at 340800 nm using a 5 nm step size. Save data.

9. Scan remaining wells (A2A7) and save data.

10. For bovine IgG at optimal wavelength, select Endpoint protocol, 96-well format, and single wavelength reading at 590 nm.

11. Highlight the appropriate wells to be read; i.e., A1 to C7 for the concentration range from 62.5 g/ml to 1 mg/ml and the blank. 12. Read absorbance. Save and analyze data.


Results
The most concentrated protein sample shows a higher peak at 590 nm than the least concentrated protein samples (Figure 1). The least concentrated samples and the blank showed higher peaks at 470 nm, whereas the more concentrated protein samples showed similar peak heights at around 470 and 590 nm. From the figure, one can conclude that whereas a peak at 470 nm is apparently caused by the cell culture medium used here, the optimal wavelength for bovine IgG is at 590 nm.

Further reading of bovine IgG concentration at 590 nm on Benchmark Plus yields a linear graph with a correlation coefficient of 0.9993 (Figure 2).


Conclusion
The Benchmark Plus Scan Well feature is used to ascertain the best wavelength for reading a desired assay. It allows scanning of a well from 340800 nm at various wavelength increments. A graph and data of the scan area are provided and can be exported into Microsoft Excel software for furt
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