What is the Pierce BCA protein assay?
Pierce BCA Protein Assay Kit is a two-component, high-precision, detergent-compatible protein assay for determination of protein concentration. Pierce BCA reagents provide accurate determination of protein concentration with most sample types encountered in protein research.
What are the steps of BCA protein assay?
Assay
- Prepare samples containing 0.2 to 50 micrograms protein in microliters.
- Add 1 ml SWR to each 20 microliters sample and mix. Incubate 30 min. at 60 degrees C.
- Cool the samples and read at 562 nm. Color will be stable for at least one hour.
What is BCA Reagent A?
BCA Protein Assay Reagent A is a component of the Pierce BCA Protein Assay Kit, a two-component, high-precision, detergent-compatible assay that is used for total protein concentration determination compared to a protein standard.
What is the difference between BCA and Bradford assay?
Historically, the BCA method is more sensitive than the Bradford method, because the first method is based on protein-copper chelation and secondary detection of the reduced copper. Whereas the Bradford method is based on protein-dye binding and colour shift from 465 to 595 nm.
How long do BCA standards last?
12 months
The BCA and Copper Reagents are stable at room temperature. The BSA Standard should be aliquoted after the first thaw and stored at -20°C. All reagents are stable for up to 12 months under proper storage conditions.
What are protein assays used for?
The purpose of the protein assay is to determine the amount or concentration of a specific protein or an array of different proteins in a sample. Isolating and detecting protein is used for many clinical and research processes.
What is the principle of the BCA assay?
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.
Is BCA light sensitive?
Protect from light: It is recommended to keep reagents within the kit box to minimize exposure to light. Directions for Use: Please see attached protocol. Background: Bicinchoninic Acid (BCA) is capable of forming an intense purple complex with cuprous ion, Cu1+, in an alkaline environment.
How long does it take to do a BCA assay?
Choose the right BCA protein assay for your sample
| Rapid Gold BCA Protein Assay | Micro BCA Protein Assay | |
|---|---|---|
| Incubation time and temperature | 5 min at RT | 60 min at 60°C |
| Total assay time | 50 min | 130 min |
| Absorbance | 480 nm | 562 nm |
| Compatible reagents | Detergents | Detergents |
Can you freeze BCA standards?
No need to freeze if for storage. The freeze-thaw cycles may actually degrade the standard protein. The data-sheet from Pierce says to store at room temp as well. After we crack open the glass vial of protein standard, we place the rest into a clean centrifuge tube and keep it stored at room temp.
Why is the BCA method used for the protein quantitative analysis?
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.
Why is BCA assay used?
BCA (Bicinchoninic Acid) Protein Assay. 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 does BCA assay stand for?
Bicinchoninic acid
Bicinchoninic acid (BCA) assay or Smith assay is a copper-based colorimetric assay for total protein quantification.
How accurate is BCA assay?
The BCA assay is not perfect. Here are some limitations that you may need to consider. The reaction may be less sensitive to the type of amino acids present in the solution but the reaction is influenced by cysteine, tyrosine and tryptophan residues.
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.
Why Pierce BCA reagents?
Used in more labs than any other detergent-compatible protein assay, Pierce BCA Reagents provide accurate determination of protein concentration with most sample types encountered in protein research.
What is the delivery information for Pierce BCA™ protein assay kits?
Delivery information: Each Pierce BCA™ protein assay kit contains BCA Reagent A (500 mL or 2×500 mL), BCA Reagent B (25 mL), and Albumin Standard Ampoules (2 mg/mL, 10×1 mL). We are currently unable to calculate your contract price for this item.
How does the BCA protein assay work?
The BCA Protein Assay combines the well-known reduction of Cu 2+ to Cu 1+ by protein in an alkaline medium with the highly sensitive and selective colorimetric detection of the cuprous cation (Cu 1+) by bicinchoninic acid (BCA). The first step is the chelation of copper with protein in an alkaline environment to form a light blue complex.
What is bicinchoninic acid protein assay?
Pierce BCA™ (bicinchoninic acid) reagents provide accurate determination of protein concentration with most sample types encountered in protein research. The Pierce BCA™ Protein 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.