BCA Quantification Kit (Broad Range)

GRiSP´s BCA Quantification Kit (Broad Range) is formulated for the quantification of total protein concentration ranging from 20-2000µg/ml. It is suitable for usage with standard spectrophotometers as well as with microplate readers.

GENERAL INFORMATION:The Bicinchoninic Acid (BCA) protein assay for the quantification of proteins is a highly sensitive colorimetric assay that is compatible with detergent solubilized protein solutions. Similar to the Lowry method, the principle is based on the Biuret reaction in which Cu2+-protein complexes are formed under alkaline condition. As proteins react as reducing agents, Cu2+ is reduced to Cu+, which is subsequently chelated by two BCA molecules, forming a purple complex that exhibits strong absorption at 562 nm. As the amount of Cu+ is proportional to the amount of protein, measuring absorption can be used to quantify protein concentration by comparing with the absorption of protein solutions of known concentrations. In comparison with the Lowry method, the BCA method is not only easier and faster, but also suffers less interference from non-ionic detergents and salts. It is insensitive to the presence of detergents such as SDS (1%) and Triton X-100.
GRiSP´s BCA Quantification Kit (Broad Range) is formulated for the quantification of total protein concentration ranging from 20-2000µg/ml. It is suitable for usage with standard spectrophotometers as well as with microplate readers. The whole procedure can be carried out in less than an hour.

KIT CHARACTERISTICS:
GRiSP´s BCA Quantification Kit (Broad Range) is compatible with the following detergents up to the given concentrations: 5% Brij®-35, 1% Brij®-56, 1% Brij®-58, 5% CHAPS, 5% CHAPSO, 5% Deoxycholic acid, 5% Nonidet P-40 (NP-40), 5% Octyl ß-glucoside, 5% Octyl ß-thioglucopyranoside, 5% SDS, 1% Span® 20, 5% Triton® X-100, 1% Triton® X-114, 1% Triton® X-305, 1% Triton® X-405, 5% Tween®-20, 5% Tween®-60, 5% Tween®-80 and 1% Zwittergent® 3-14.

It is also not interfered by the following reducing and thiol containing agents up to the given concentrations: 10% N-acetylglucosamine (in PBS), 1mM DTE, 1mM DTT, 10mM Glucose, 0.01 ß-mercaptoethanol, 3M Potassium thiocyanate and 0.01% Thimerosal. Moreover, the presence of chelating agents EDTA up to 10mM and Sodium Citrate (up to 200mM) have no negative effect on the outcome of the assay.

In general, chelating reagents or reagents that change the pH or reduce copper are known to interfere with the assay. These include ascorbic acid, cathecholamines, creatinine, cysteine, EGTA, glycerol (impure), hydrogen peroxide, hydrazides, iron, lipids, melibiose, phenol red, sucrose (impure), tryptophan, tyrosine and uric acid.