Validating the specifications, value and safety of your raw materials, products and assets. Formally confirming that your products and services meet all trusted external and internal standards. Protein quantification or total protein content by colorimetric methods such as absorbance at , Bradford or BCA assays and quantitative amino acid analysis.
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.
Careful selection of the protein quantification method is required to ensure accurate data. Colorimetric assays, for example, are dependent on the use of an external reference standard and the different absorbance properties between product and the reference standard may lead to inaccuracies.
Quantitative amino acid analysis can also be used for protein content determination but an understanding of how different proteins hydrolyze under similar conditions and the stability of amino acids during hydrolysis is critical. Our scientists are experts in the selection of protein quantification assays which is underpinned by a deep knowledge of the physicochemical properties of proteins and amino acids.
We address any potential source of variability in the accuracy of measurement of total protein content, delivering precise and robust data every time. Colorimetric assays such as the Bradford or bicinchoninic acid BCA assays measure UV-light absorbance and can be used to calculate protein concentration from the absorbance measurement, once the extinction coefficient molar absorptivity has been accurately established.
Other assays include absorbance at nm, Lowry assay and the Biuret assay. Protein quantification can be offered from our cGMP compliant laboratories as a standalone test or as part of a biologic release testing package for commercial release or a biopharmaceutical stability study test.
As your outsourcing partner for biologics or protein product development, our expertise in protein content can accelerate development timelines through delivering accurate and robust analytical services. Bringing quality and safety to life, we offer Total Quality Assurance expertise to help you to meet and exceed quality, safety and regulatory standards.
View our comprehensive collection of digital resources. The Bradford assay is another colorimetric assay that is compatible with most buffers, reagents and preparations. It is relatively easy to use since it requires no working reagent preparation and is proven to give fast, accurate results. This is recommended for determining protein content of cell fractions and for assessing protein concentrations prior to conducting gel electrophoresis.
Since there is no single assay that is sensitive to all protein types, you need to have a working knowledge on how each assay works so you can choose the most suitable one to use for each of your applications. Image By: Joe Dunckley. Topics: Protein Estimation , Protein Concentration. This assay is suitable for the simple and rapid estimation of protein concentration. This assay is based on a single Coomassie dye based reagent. The binding of protein to the dye results in a change of color from brown to blue.
The change in color density is proportional to protein concentration. These aptamers are created and selected for using SELEX technology , which is an adaptation of dynamic combinatorial chemistry techniques.
The use of oligonucleotides in place of antibodies opens up a new realm of conjugation and material possibilities for protein assay. Related news. He has conducted research at German and Spanish academic institutions. Having worked in companies in Ireland, USA and Spain, he has more than 20 years of experience applying magnetic materials and sensors to industrial products and processes.
He has filed several international patents on the field and co-authored more than 20 scientific papers, most of them on the subject of magnetic particle movement. Veritas Corporation Tel. Zenmindes Biotechnology Tel. Posted on Thu, Jul 22, Purpose of Protein Assays T he 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.
Protein assay types 1. UV absorbance and spectrophotometry : The aromatic side chains of amino acids absorb light in the UV range. HPLC high-performance liquid chromatography The sample is carried by liquid solvent at high pressure through an adsorbent column. Different amino acid side chains interact more or less with the material. The side chains that react the least move fastest through the column and those that react the most move slowly.
The proteins are fractionated as they flow out of the column. You can read more about immunoaffinity chromatography in our article " immunoaffinity chromatography ". A standard curve is used to determine protein amount.
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