Measure Protein BCA
88 NanoDrop One User Guide Thermo Scientific
Protein assay kits and protocols
Please refer to the NanoDrop website for up-to-date kits and protocols for the
NanoDrop One instruments. Follow the assay kit manufacturer’s recommendations for all
standards and samples (unknowns). Ensure each is subjected to the same timing and
temperature throughout the assay.
Protein standards for generating a standard curve may also be provided by the kit
manufacturer. Since the NanoDrop One pedestals can measure higher protein concentrations
than traditional cuvette-based spectrophotometers, you may need to supply your own protein
standards at higher concentrations than provided by the manufacturer. For example,
additional standards may be required to ensure the standard curve covers the dynamic range
of the assay and the expected range of the unknown samples.
Working with standard curves
A standard curve is required for colorimetric protein analysis.
• Each experiment requires a new standard curve.
• Prepare standards and unknown samples the same way. See the kit manufacturer’s
guidelines and recommendations.
– All reference and standards solutions should be the same buffer used to resuspend
the samples plus the same volume of reagent added to the samples.
– First standard is a reference measurement. The reference solution should contain
none of the analyte of interest. (The reference measurement is not the same as a blank
measurement. This application requires both.)
– Concentration range of the standards must cover the dynamic range of the assay
and the expected range of the unknown samples. Sample analyte concentrations are
not extrapolated beyond the concentration of the highest standard.
• Use the application setup screen to enter concentration values for the standards and to
specify how standards and samples will be measured (number of replicates, etc.).
– Depending on the Curve Type setting, a standard curve can be generated using two
or more standards.
–The software requires one reference measurement and allows up to 7 standards.
– Concentration values for standards can be entered in any order but the standards
must be measured in the order in which they were entered; however, best practice
dictates that standards be measured from the lowest concentration of the standard
analyte stock to the highest.
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