EXP 2.docx - BIO 462 – BIOCHEMISTRY Experiment 2 Protein Determination Group AS2463C Lecturer’s Name Dr Lyena Watty Zuraine Binti Ahmad Group

EXP 2.docx - BIO 462 – BIOCHEMISTRY Experiment 2 Protein...

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BIO 462 – BIOCHEMISTRY Experiment 2: Protein Determination Group: AS2463C Lecturer’s Name: Dr. Lyena Watty Zuraine Binti Ahmad Group Members Matric ID Aiman Nabilah Binti Kamarulazmee 2019722673 Fariha Natasha Binti Faijal 2019725877 Inshirah Binti Izham 2019316995 Nur Allia Syazana Binti Mohd Shah 2019725501 Zaiti Nadira Binti Md Khalid 2019555081 Date of Submission: 07 October 2019 INTRODUCTION Every chemical compound absorbs, transmits, or reflects light over a certain range of wavelength. Spectrophotometry is a measurement of how much a chemical substance
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absorbs or transmits. It is widely used for quantitative analysis in various areas such as in chemistry, physics , and any application that deals with chemical substances or materials can use this technique. In biochemistry, for example, it is used to determine enzyme-catalyzed reactions. In clinical applications, it is used to examine blood or tissues for clinical diagnosis. There are also several variations of the spectrophotometry such as atomic absorption spectrophotometry and atomic emission spectrophotometry (Grimsley et. al, 2003). Next, a spectrophotometer is an instrument that measures the amount of intensity of the light absorbed after it passes through sample solution. The amount of a known chemical substance can also be determined by measuring the intensity of light detected using spectrophotometer. Spectrophotometer can be classified into two different types depending on the range of wavelength of the light source. For example, UV-visible spectrophotometer uses light over the ultraviolet range (185 - 400 nm), while IR spectrophotometer uses light over the infrared range (700 - 15000 nm) of electromagnetic radiation spectrum. In the experiment to determine the protein concentration, Biuret assay, Bradford assay and Lowry assay are used. The biuret test is a chemical test used for detecting the presence of peptide bonds . The principle of biuret test is conveniently used to detect the presence of proteins in biological fluids. While, Biradford assay is a rapid and accurate method for the estimation of protein concentration is essential in various areas of biology and biochemistry. An assay originally described by Bradford has become the preferred method for quantifying protein in many laboratories. This technique is simpler, faster, and more sensitive than the Lowry method. Moreover, when compared with the Lowry method, it is subject to less interference by common reagents and non-protein components of biological samples ( Jones, 2007). Meanwhile, Lowry protein assay is a biochemical assay for determining the total level of protein in a solution total protein concentration is exhibited by a color change of the sample solution in proportion to protein concentration, which can then be measured using colorimetric techniques. The method combines the reactions of copper ions with the peptide bonds
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