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Protein concentration Essay Example

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Protein concentration

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The blank test tube will be added with 0.20ml 85% Sodium chloride Solution. The blank test tube will be used as reference test tube. This will be the test tube for comparison purposes with the other test tubes containing the various reagents. The blank test tube will be the first tube to be read in the spectrophotometer that will set back the reading to zero. All the other test tubes would be read in comparison with the blank test tube’s reading to determine significant results and readings.The rest of the test tubes would be added with certain volume of reagents such as Biuret reagent and Folin and Ciucalteu’s reagent, mixed and allowed to stand for different holding times. The contents of the test tubes would then be read with the absorbance with the blank solution as reference. The protein concentration of every diluted sample would be determined.The use of Biuret Method is greatly used for protein analysis. Biuret is a compound formed from combination of urea through heat producing amide groups. A peptide bond results with the amide group of the biuret reagent and the carboxyl group. This would result to blue color solution observable through spectrophotometer in the presence of copper ion complex (Biuret Protein Assay, 2010). The assay would greatly result to colored solution in alkali environment (Detection and Assay Methods, 1995).One of the reagents used in the experiment is the Folin and Ciocalteu’s Phenol reagent. This mixture of phosphomolybdate and phosphototungstate (MP Biomedicals, 2014) is useful a reagent commonly used for portein concentration determination through the Lowry method. The addition of this reagent to a previously pretreated diluted sample generates chromogens that increase absorbance to 550-750 nm. Color intensity is affected by the presence of tryphtophan and tyrosine (Sigma-Aldrich, 2014). Under alkaline conditions, a monovalent ion and radical groups of tryhtophan and tyrosine reacts with the Folin reagent turning the solution into tungsten blue that can be viewed on spectrophotometer (Caprette, 2012).Based on the results it can be observed that the absorbance and protein concentration is linear in nature. With the use of various reagents, it can be deduced that the methodology used in getting data for protein content can be translated into graphical representation. It can be seen through the

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Works Cited

Anthis and Clore. (2013). Sequence-specific determination of protein and peptide concentrations

by absorbance at 205nm. National Center for Biotechnology Information, US National

Library of Medicine.

Biuret Protein Assay. (2010). Gru.edu. Accesible through

www.gru.edu/colleges/scimath/biology/courses/.../biuretproteinassay.pdf‎

Craic Technologies. (2012). Ultraviolet Absorbance of Protein. Craic Technologies, Inc.

Caprette, D. (2012). Hartree-Lowry and Modified Lowry Protein Assays. Rice University

Detection and Assay Methods. (1995). Assays for Total Protein. John Wiley & Sons. Inc.

Knight and Chambers.(2003). Problems Associated with Determining Protein Concentration.

Molecular Biotechnology . Accessible through

http://link.springer.com/article/10.1385/MB%3A23%3A1%3A19#page-1

MP Biomedicals. (2014). Folin & Ciocalteu’s Phenol Reagent. MP Biomedicals LLC.

Accessible through http://www.mpbio.com/product.php?pid=02195186&country=168

Pierce. (2014). Protein Assay Technical Handbook. Pierce Biotechnology.

Pierce Protein Biology Products. (2014). Methods for Detecting Protein-DNA Interactions.

Thermo Fisher Scientific.

Reusch, William. (2013). “Protein, Peptides, And Amino Acids. Accessed through

https://www2.chemistry.msu.edu/faculty/reusch/virttxtjml/proteins.htm

Sapan, et al. (1999). Colorimetric Protein Assay Techniques. Biotechnol Appl Biochem. US

National Library of Medicine National Institutes of Health.

Science Learning Hub. (2014). Protein. The University of Waikato.

Sigma-Aldrich. (2014). Folin & Ciocalteu’s Phenol Reagent. Sigma-Aldrich Co. Accessible through http://www.sigmaaldrich.com/catalog/product/sial/f9252?lang=en&region=PH

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