Thermo Scientific™ Pierce™ Dilution-Free™ Rapid Gold BCA Protein Assay

Description

• The Pierce Dilution-Free Rapid Gold BCA Protein Assay Kit is an effortless, fast, and accurate assay optimized to determine total protein concentration
• It is detergent-compatible and has a 5X wider dynamic range than traditional protein assays of 0.02–10 mg/mL
• This wide dynamic range eliminates the need to dilute most samples to get them into range of the assay
• Additionally, the Dilution-Free Rapid Gold BCA Protein Assay comes with a set of seven pre-diluted BSA protein standards packaged in a multichannel pipette-friendly tubestrip, so no serial dilutions are required
• The tubestrip includes a single empty tube that enables users to add their own sample buffer for the purpose of blank subtraction
• The Dilution-Free Tubestrip Holder (Cat
• No
• A57184) enables easy handling on the bench
• Features of the Dilution-Free Rapid Gold BCA Protein Assay Kit include: Dilution-free —eliminates the need for sample and standard curve dilutions, reducing set-up time by up to 80% Short assay time —five-minute, room temperature reaction Wide assay range —linear working range for BSA of 20 to 10,000 μg/mL Uniform —exhibits similar protein-to-protein variation as traditional BCA Protein Assay and less protein-to-protein variation than dye-binding protein assay methods (Bradford) Compatible —unaffected by typical concentrations of most ionic and non-ionic detergents Colorimetric —optimal absorbance at 480 nm Accurate —2X more accurate than Bradford assays The Pierce Dilution-Free Rapid Gold BCA Protein Assay Kit uses the same copper-chelating technology as the well-known traditional Pierce BCA Protein Assay and provides comparable accuracy, but with a five-minute, room temperature incubation and measured at 480 nm
• This improvement eliminates the need to perform serial or sample dilutions, endure a lengthy incubation period, or expose the samples to elevated temperatures yielding faster time to results
• The traditional BCA Protein Assay is a widely used and accepted protein assay, trusted for its highly accurate protein concentration determination and compatibility with most sample types encountered in protein research
• However, in order to develop the samples in this traditional protocol, one must either heat the reaction at 37°C for 30 minutes or allow room temperature development for two hours
• The Dilution-Free Rapid Gold BCA Protein Assay maintains the key characteristics of the traditional BCA assay but allows a fast time and room temperature incubation equal to dye-binding methods
• The Dilution-Free Rapid Gold BCA Assay can be used to assess yields in whole cell lysates and affinity-column fractions, as well as to monitor protein contamination in industrial applications
• Similar to the traditional BCA assay, the Dilution-Free Rapid Gold BCA Assay is affected much less than dye-binding methods by protein compositional differences, providing low protein-to-protein variation
• How the Dilution-Free Rapid Gold BCA Protein Assay detects protein The Dilution-Free Rapid Gold BCA Protein Assay uses the same copper reduction method as the traditional BCA Protein Assay with a unique copper chelator
• The assay combines the well-known reduction of Cu2+ to Cu1+ by protein in an alkaline medium with the highly sensitive and selective colorimetric detection of the cuprous cation (Cu1+) by the proprietary chelator
• The first step is the chelation of copper with protein in an alkaline environment to form a light green complex
• In this reaction, known as the biuret reaction, peptides containing three or more amino acid residues form a colored chelate complex with cupric ions in an alkaline environment containing sodium potassium tartrate
• In the second step of the color development reaction, the Dilution-Free Rapid Gold BCA chelator reacts with the reduced (cuprous) cation that was formed in step one
• The intense gold-colored reaction product results from the chelation of two molecules of BCA with one cuprous ion
• The BCA/copper complex is water soluble and exhibits a strong linear absorbance at 480 nm with increasing protein concentrations
• The complex is approximately 100 times more sensitive (lower limit of detection) than the pale green color of the first reaction
• The reaction that leads to the color formation is strongly influenced by four amino acid residues (cysteine, cystine, tyrosine, and tryptophan) in the amino acid sequence of the protein
• However, unlike the Coomassie dye-binding methods (i.e
• Bradford), the universal peptide backbone also contributes to color formation, helping to minimize variability caused by protein compositional differences
• Compatibility of the Dilution-Free Rapid Gold BCA Protein Assay Certain substances are known to interfere with the Dilution-Free Rapid Gold BCA Protein Assay, including those with reducing potential, chelating agents, and strong acids or bases
• Because they are known to interfere with protein estimation at even minute concentrations, avoid the following substances as components of the sample buffer: Ascorbic acid EGTA Iron Impure sucrose Catecholamines Impure glycerol Lipids Tryptophan Creatinine Hydrogen peroxide Melibiose Tyrosine Cysteine Hydrazides Phenol red Uric acid TCEP Thimerosal 2-mercaptoethanol Pierce Dilution-Free Tubestrip Holder The Pierce Dilution-Free Tubestrip Holder is a reusable foam stand for use with the Pierce Dilution-Free BSA Protein Standards and is included with the 20 mL trial size of the Pierce Dilution-Free Rapid Gold BCA Protein Assay Kit
• It is also available for sale separately
• Protein standard concentrations are printed on the Tubestrip Holder for easy reference and pipetting on the benchtop.