Measure Protein Bradford
102 NanoDrop One User Guide Thermo Scientific
To maximize reliability with the Protein Bradford assay:
• Work quickly and do not allow prepared standards or samples to sit longer than
necessary. Coomassie dye-dye and Coomassie dye-protein aggregates can form
particulates with increasing development time, resulting in significant fluctuations in
absorbance readings.
• Measure standards and samples in triplicate using a new aliquot for each measurement.
For pedestal measurements, the total analyte (protein-dye) signal at 595 nm is limited to
~0-0.150A due to the pedestal’s 1.0 mm pathlength, the Coomassie dye concentration,
and the acidic pH.
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.
To measure Protein Bradford standards and samples
Note If you have a NanoDrop One
C
model instrument, using the cuvette option will
result in a higher absorbance signal.
NOTICE
• Do not use a squirt or spray bottle on or near the instrument as liquids will flow into
the instrument and may cause permanent damage.
• Do not use hydrofluoric acid (HF) on the pedestals. Fluoride ions will permanently
damage the quartz fiber optic cables.
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