What is BCA protein assay protocol?
The BCA protein assay is used for quantitation of total protein in a sample. The principle of this method is that proteins can reduce Cu+2 to Cu+1 in an alkaline solution (the biuret reaction) and result in a purple color formation by bicinchoninic acid.
What is Micro BCA?
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). The assay is exceptionally linear and exhibits very low levels of protein-to-protein variability.
How does the Pierce BCA protein assay work?
The Pierce BCA Protein Assays combines the well-known reduction of Cu2+ to Cu+ by protein in an alkaline medium (also known as the biuret reaction) with the highly sensitive and selective colorimetric detection of the cuprous cation (Cu+) by bicinchoninic acid (BCA).
How long does it take to do a BCA assay?
BCA Standard Protocol assay was performed on the Modulus using the Absorbance Module and the 560 nm filter. Following a 37°C incubation for 30 minutes, the samples cooled to room temperature before analysis on the Modulus.
How long does a BCA take?
Typically the process can be completed in less than six weeks. It may be shorter depending on current volume, how long it takes vendors to complete and submit forms/contracts, and whether the vendor employee would have logical (ex.
Why is BSA used in BCA assay?
The intensity of the color formed by these assays is measured by absorbance photometry at 595 nm and 562 nm for the Bradford and BCA assays, respectively. Typically, standard solutions of bovine serum albumin (BSA) are used to produce a calibration curve of absorbance versus mass concentration.
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 the principle of BCA?
The principle of the bicinchoninic acid (BCA) protein assay1 relies on the formation of a Cu2+-protein complex under alkaline conditions, followed by reduction of the Cu2+ to Cu1+. The amount of reduction is proportional to the amount of protein present.
Does blood interfere with BCA assay?
All Answers (3) Hi Chris, hemoglobin absorbs light around the same wavelength used for the BCA assay. So yes, presence of RBCs (hemoglobin) could interfere with the accuracy of the assay.
Which protein assay is most sensitive?
Compared to other methods, the BCA assay is one of the most sensitive (it can detect proteins at concentrations as low as 5 ug/mL). It has less variability than others (i.e., Bradford assay), and it can be used to measure a wide range of protein concentration.
How do you dilute a BCA sample?
This is how I typically dilute my samples:
- Transfer 20 µL of each sample to a new cDNA tube/Eppendorf tubes.
- Add 60 µL of deionized water to the samples (dilute by a factor of 4).
How does the micro BCA protein assay reagent kit work?
The Micro BCA™ Protein Assay Reagent Kit uses concentrated reagents and a protocol that utilizes an extended incubation time at an elevated temperature (60°C). The result is an extremely sensitive colorimetric protein assay in a test tube or microplate assay format.
Which is Thermo Scientific Pierce BCA protein assay kit?
The Thermo Scientific Pierce BCA Protein Assay Kit is a two-component, high-precision, detergent-compatible assay reagent set to measure (A562nm) total protein concentration compared to a protein standard. Features of the BCA Protein Assay Kit: • Colorimetric—estimate visually or measure with a standard spectrophotometer or plate reader (562nm)
Which is the best protein quantitation assay kit?
Specifications Assay: Protein Quantitation Assay Product Line: Micro BCA™, Pierce™ Description: Pierce Micro BCA Protein Assay Kit For Use With (Application): Solution-based Detection, Absorbance Specificity: Not Target-Specific
Is the BCA protein assay sensitive to cuprous cation?
The BCA Protein 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 bicinchoninic acid.