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The Diagnostic Uses of Electrophoresis
The Diagnostic Uses of Electrophoresis
The Diagnostic Uses of Electrophoresis
PRESENTATION OUTLINE
ELECTROPHORESIS
CONCLUSION
REFERENCES
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ELECTROPHORESIS
INTRODUCTION Electrophoresis is a technique for separation of different charged particles.
It is based on movement of charged particles through a solution
PRINCIPLE In an electrophoresis system, particles take on electrical charge, move toward either the cathode(negative electrode) or the anode(positive electrode).
Separation is based on the differences in their charge-to3
CONVENTIONAL ELECTROPHORESIS
SET-UP
4.
Two buffer boxes containing the buffer solution Electrodes connected to the power supply. The electrophoresis support which contacts the buffer by means of the wicks. Wicks
BUFFERS The buffer carries the applied current and also determines the electrical charge on the particles.
SUPPORT MEDIA The support medium provides the matrix on which separation takes place. Types of support media include: starch, cellulose acetate, agarose and polyacrylamide gels. CLINICAL APPLICATIONS To extensively study families of proteins in order to detect 5 genetic- or disease-based differences;
DETECTION AND QUANTIFICATION The support medium is placed in a densitometer in which the gel (or other support medium) is moved past a measuring optical system;
The absorbance of each fraction is displayed on a
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(Johnson, 2005)
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(Johnson, 2005)
Synthesis of the acute-phase proteins is a direct result inflammation triggered by tissue damage of any kind.
o CHRONIC INFLAMMATORY STATES A diffuse rise in -globulin coupled with the usual increase in the -1 and -2 fractions of the acute phase. 13
Albumin and alpha1-globulin: reduced, -globulin: markedly increased, with apparent fusion of the - and bands. o NEPHROTIC SYNDROME Reduced albumin, alpha1 and sometimes -globulin; Increase in alpha2-globulin due to a relative increase in 2macroglobulin.
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DEFICIENCY:
The 1-band consists almost entirely of AAT : absence or an obvious reduction in the density strongly suggests AAT deficiency.
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CONCLUSION
It is recommended that full use of this diagnostic technique be made for better diagnostic sensitivity and
specificity.
In addition, correct monitoring of buffer pH,
voltage, equal
quantity of test and control applied and skill of
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REFERENCES
Berg JM, Tymoczko JL Stryer L (2002).
Chemistry and Metabolic Medicine, 7th ed. London: Edward Arnold, 2006:280-300.
Johnson AM. Amino acids, peptides and proteins.
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In: Burtis CA, Ashwood ER, Bruns DE, eds. Tietz textbook of clinical chemistry and molecular
REFERENCES (CONTINUED)
Karcher RE, Landers JP. Electrophoresis. In:
Burtis CA, Ashwood ER, Bruns DE, eds: Tietz textbook of clinical chemistry and molecular diagnostics, 4th ed. Philadelphia: Saunders, 2006:121-40.
Landers JP. Molecular diagnostics on
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