Thermo Scientific NanoDrop One User Guide 87
Measure Protein BCA
Measure Total Protein Concentration
The Protein BCA assay uses bicinchoninic acid as a colorimetric detection reagent to
determine total protein concentration in unpurified protein samples. This application is
useful for measuring dilute protein solutions or proteins in the presence of components that
exhibit significant absorbance between 200 nm and 280 nm, which rules out direct protein
measurements at 280 nm or 205 nm. This application measures absorbance at 562 nm and
uses a standard curve to calculate protein concentration. A single-point baseline correction is
applied.
Theory of Protein BCA assay
The Protein BCA assay uses bicinchoninic acid (BCA) as the detection reagent for Cu
+1
,
which is formed when Cu
+2
is reduced by certain proteins in an alkaline environment. A
purple reaction product is formed by the chelation of two molecules of BCA with one cuprous
ion (Cu
+1
). The resulting Cu-BCA chelate formed in the presence of protein is measured at
562 nm and baseline-corrected using the absorbance value at 750 nm. Pre-formulated kits of
BCA reagent and CuSO
4
are available from us or a local distributor.
Measures total protein
concentration of unpurified protein
samples using a bicinchoninic acid
colorimetric detection reagent.
Measure Total Protein
Reported Results
Settings
Detection Limits
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