What is the Pierce BCA protein assay based on?
bicinchoninic acid
Discard any kit reagent that shows discoloration or evidence of microbial contamination. The Thermo Scientific™ Pierce™ BCA Protein Assay is a detergent-compatible formulation based on bicinchoninic acid (BCA) for the colorimetric detection and quantitation of total protein.
What does the BCA assay measure?
The BCA protein assay is used to quantify total protein in a biological sample. BCA stands for Bicinchoninic acid, which is the key reagent used to produce a colored product. The purple colored product is analyzed in reference to a standard curve in order to quantify protein concentration.
What is the diluent for BCA assay?
BSA is hydrophobic, however it dissolved well in slightly luke warm distilled water or PBS (pH 7.2). Use odd series of concentration of BSA say 10, 30, 50, 70, 90 and 110 microgram/ml to calibrate a reference profile of BSA.
What is Microbca?
The Pierce Micro BCA Protein Assay Kit is a three-component version of our BCA reagents, optimized to measure total protein concentration of dilute protein solutions (0.5 to 20 µg/mL).
Is BCA reagent light sensitive?
It is recommended that the BCA Compatibility Reagent be used to decrease interference from reducing agents, chelators, detergents, and other common ingredients found in most lysis buffers. Protect from light: It is recommended to keep reagents within the kit box to minimize exposure to light.
Does urea interfere with BCA?
But remember to prepare standards in the same buffer conditions. Manyak’s solution will be likely also work, so give a try as BCA assay is so common that every lab has the reactants. Use the RcDc Protein Assay (Bio-Rad). It is compatible with detergents and urea.
How does a BCA work?
BCA rely on the formation of a Cu2 +–protein complex in a basic environment, followed by reduction of the Cu2 + to Cu+ (Smith et al., 1985). The amount of Cu2 + that is reduced is proportional to the amount of protein present in solution.
How does BCA assay calculate protein concentration?
You can use this formula to calculate the amount of protein in your sample: [ (OD of the test sample) / (OD of the standard sample) ] X concentration of the standard sample. = 4.0 mg/mL protein (in test sample).
How do you make BSA dilutions?
Mix 50 μl of the BSA standard solution (2 mg/ml) with 950 μl of diluent and mix well to prepare a 0.1 mg/ml BSA standard solution. Prepare 2 sets of the dilutions of BSA standard solution as shown below 1.5 ml microtubes (7 types x 2 sets = 14 microtubes).
How do you dilute BCA standards?
A. Dilute the contents of one Albumin Standard (BSA) ampule into several clean vials, preferably using the same diluent as the sample(s). Each 1 ml ampule of 2.0 mg/ml Albumin Standard is sufficient to prepare a set of diluted standards for either working range suggested in Table 1.
What is the difference between BCA and Micro BCA assay?
While both assays use BCA (bicinchoninic acid) to measure protein concentration of dilute protein samples, the Micro BCA assay is designed for testing proteins in lower sample concentrations of 0.5-20 ug/ml, while the standard BCA assay works in the range of 20-2000 ug/ml.
What is BCA chemical?
Bicinchoninic acid /baɪsɪnkɔːnɪnɪk/ is a weak acid composed of two carboxylated quinoline rings. Bicinchoninic acid is most commonly employed in the bicinchoninic acid (BCA) assay, which is used to determine the total level of protein in a solution.