Basis of differentiation | ||
Definition | Is a spectroscopic method technique for measuring the concentration of protein in a solution | Is a lab technique used to measure the total amount of protein in a sample |
Accuracy | More accurate | Less accurate |
Time | Is relatively fast | Is comparatively more time-consuming |
Sensitivity | Is typically more sensitive, capable of detecting protein concentrations as low as 1-20 µg/mL | Is slightly less sensitive, typically detecting protein concentrations in the range of 25-2000 µg/mL |
Mechanism | Based on the absorbance shift of Coomassie Brilliant Blue G-250. In its unbound form, this dye is reddish-brown. When the dye binds to proteins, it undergoes a color change from reddish-brown to blue. | Relies on the reduction of cupric ions (Cu²⁺) to cuprous ions (Cu⁺) in the first reaction. In the second reaction, BCA reacts with the reduced cuprous ions (Cu⁺). This reaction forms a stable, deep purple complex. |
Compatibility with agents | Is less tolerant of detergents and other chemical additives, with high levels of detergents, reducing agents and buffers, potentially impacting the results | Is more resistant to a broad range of detergents, reducing agents and buffers because of its two-step reaction process |