Agarose electrophoresis of serum proteins and immunofixation of monoclonal proteins

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The identification and quantification of proteins in body fluids generally provides essential information on the pathologic condition of an individual (Bossuyt et al. , 1998). One of the most commonly examined body fluids is serum, which is a component of blood that is collected after the removal of blood cells and fibrin clots (Jenkins & Guerin, 1995). Despite the water-like features of serum, this fluid contains a number of proteins that assist healthcare professionals in determining the actual medical disorder that a patient may have developed (Van Duyne et al. , 2010).

In order to analyze the amounts of various proteins in serum, there is a need to employ a sensitive and reliable technique that allows the separation of each protein from the fluid sample. This laboratory report will present the results of an experiment that employed agarose gel electrophoresis in separating proteins from serum. This experiment aims to familiar the student with the concept of protein separation using a specific biological sample, namely serum, using a technique known as agarose gel electrophoresis.

In addition, the experiment aims to perform protein analysis based on the bands that are generated from the electrophoretic separation of serum proteins. METHODOLOGY The experiment was performed according to the description provided in Session 2 of the Clinical Biochemistry 3 Laboratory Manual 2010, pages 4 to 9. Briefly, 4 cases were loaded onto a QuickGel Split Beta SPE Gel, together with two controls, SPE Normal Control and SPE Abnormal Control for reference (Jimenez et al. , 1995).

An electric field was then introduced to the gel to allow migration of proteins through the substrate by applying approximately 300 volts for 12 minutes. The gel was thereafter stained and washed using the recommended kit solutions and subsequently dried. The gel was scanned to determine the densities of each protein band in each sample. RESULTS The electrophoretic separation of serum proteins over an agarose substrate generated multiple bands of specific molecular weights. Table 1 presents the densitometric composition of proteins from the 4 serum samples, with reference to normal and abnormal controls.

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