The BCA (bicinchoninic) protein assay is a widely used method for colorimetric detection and quantitation of total protein in a solution . The BCA protein assay is a Copper-based protein assay and is also known as the Smith assay because it was introduced by Paul K.
How does a BCA protein assay work?
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 BCA used for?
The BCA (bicinchoninic) protein assay is a widely used method for colorimetric detection and quantitation of total protein in a solution . The BCA protein assay is a Copper-based protein assay and is also known as the Smith assay because it was introduced by Paul K.
What is being detected in the BCA protein assay?
In addition, the BCA Assay responds more uniformly to different proteins than the Bradford method. The BCA Protein Assay combines the protein-induced biuret reaction (see above) with the highly sensitive and selective colorimetric detection of the resulting cuprous cation (Cu1+) by bicinchoninic acid (BCA).
What type of assay is the BCA method?
3.2 Bicinchoninic Acid Protein Assay. Bicinchoninic acid (BCA) assay or Smith assay is a copper-based colorimetric assay for total protein quantification . BCA rely on the formation of a Cu 2 + –protein complex in a basic environment, followed by reduction of the Cu 2 + to Cu + (Smith et al., 1985).
How does a BCA work?
BCA rely on the formation of a Cu 2 + –protein complex in a basic environment, followed by reduction of the Cu 2 + to Cu + (Smith et al., 1985). The amount of Cu 2 + that is reduced is proportional to the amount of protein present in solution.
What does BCA 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.
How long does a BCA assay take?
| Rapid Gold BCA Protein Assay BCA Protein Assay | Incubation time and temperature 5 min at RT 30 min at 37°C | Total assay time 50 min 100 min | Absorbance 480 nm 562 nm | Compatible reagents Detergents Detergents |
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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 .
Why is BSA used in BCA assay?
BSA is used because of its stability to increase signal in assays , its lack of effect in many biochemical reactions, and its low cost, since large quantities of it can be readily purified from bovine blood, a byproduct of the cattle industry.
Why BCA protein assay is most sensitive method?
The BCA assay has a lot of advantages. 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 protein assays work?
The dye binding protein assay is based on the binding of protein molecules to Coomassie dye under acidic conditions . ... The basic amino acids, arginine, lysine and histidine play a role in the formation of dye-protein complexes color. Small proteins less than 3kDa and amino acids generally do not produce color changes.
What does BCA stand for in chemistry?
BCA stands for before-change-after . Students start by filling in the quantity of reactants that are present before the chemical reaction happens.
Which protein assay is the best?
- Bicinchoninic Acid (BCA) This colorimetric, two-step assay was originally developed in 1985 – making it a baby compared with the 64-year-old Lowry assay! ...
- Bradford. ...
- Folin-Lowry. ...
- Kjeldahl. ...
- Ultraviolet Absorption.
How accurate is Bradford assay?
The Bradford assay is very fast and uses about the same amount of protein as the Lowry assay. It is fairly accurate and samples that are out of range can be retested within minutes. ... It is sensitive to about 5 to 200 micrograms protein, depending on the dye quality.
Does SDS interfere with BCA assay?
The 2- mercaptoethanol will interfere with the BCA assay, and the SDS will interfere with the Bradford assay.
