How does NanoOrange assay work?
To perform a protein assay, the protein sample is simply added to the NanoOrange™ reagent in a specialized diluent and this mixture is heated at 95° C for ten minutes. Fluorescence can be measured as soon as the mixture has cooled to room temperature.
What is NanoOrange?
The NanoOrange Protein Quantitation Kit contains a very sensitive and easy assay for protein quantitation, with detection as low as 10 ng/mL of protein in solution. This fluorescent dye is suitable for use with spectrofluorometers and microplate readers.
What is the purpose of protein quantitation?
Protein quantification is necessary to understand the total protein content in a sample or in a formulated product. Accurate protein quantification is important as a range of other critical assays require precise total protein content results in order to generate data.
How do you calculate quantification of a protein?
The quickest way to estimate the amount of protein in solution is to use UV-vis to measure absorbance directly, but this is generally not very accurate or sensitive. Highly accurate quantitation of most proteins can be achieved using either a Bradford or bicinchoninic acid (BCA) assay.
What is protein determination?
Direct protein determination is when protein content is calculated based on the analysis of amino acid residues. Indirect protein determination can for instance be inferred following the determination of the nitrogen content, or after chemical reactions with functional groups within the protein.
Which method is best for protein estimation?
Top 5 Protein Quantification Assays
- 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!
- Ultraviolet Absorption.
What is quantitative determination of proteins?
Spectrophotometric protein quantitation assays are methods that use UV and visible spectroscopy to rapidly determine the concentration of protein, relative to a standard, or using an assigned extinction coefficient.
What is the significance of the factor 6.25 when calculating protein content?
Following the nitrogen determination, crude protein content is calculated using a conversion factor. The original, and still frequently used, conversion factor 6.25 is based on an assumption that the general nitrogen content in food proteins is 16% and that all nitrogen in foods is protein-bound.
What are the four major methods of determining protein concentration?
Due to their biochemical character, these components are analyzed using proteomic techniques such as electrophoresis, chromatography and mass spectrometry. A very important stage of such studies is the measurement of protein concentration in the sample, which is most often performed by colorimetric methods.
What are the 3 analytical methods for determining protein content?
There are three major protein analysis techniques: protein separation, western blotting and protein identification.
What’s the purpose of protein assay?
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 Kjeldahl method protein determination?
The Kjeldahl method (the industry standard test used to routinely measure the crude protein content of foods) measures the total nitrogen content of a food, which is then used to estimate the crude protein content by applying a conversion factor to the result.