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5
Protein Applications
Measure Protein Bradford
Protein assay kits and protocols
To measure Protein Bradford standards and samples
110
NanoDrop One User Guide
• 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.
Note If you have a NanoDrop One
option will result in a higher absorbance signal.
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.
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.
Before you begin...
Before taking pedestal measurements with the NanoDrop One instrument, lift the
instrument arm and clean the upper and lower pedestals. At a minimum, wipe the
pedestals with a new laboratory wipe. For more information, see
Pedestals.
To measure Protein Bradford standards and samples
1. From the Home screen, select the Proteins tab and tap Protein Bradford.
2. Specify a
curve type
concentration of each
Tip: For this assay, set Curve Type to "2nd Order Polynomial" and
Replicates to 3.
C
model instrument, using the cuvette
and number of
replicates for each standard
standard.
Cleaning the
and enter the
Thermo Scientific
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