Blood cells electrophoresis

Triose phosphate isomerase (TPI) catalyzes the interconversion of glyceralde-hyde-3-phosphate and dihydoxyacetone phosphate and has an important role in glycolysis, gluconeogenesis, fatty acid synthesis, and the hexose monophosphate pathway. Red blood cell TPI activity measured in vitro is approximately 1000 times that of Hx, the least active glycolytic enzyme. TPI is a dimer of identical subunits, each of molecular weight 27,000, and does not utilize cofactors or metal ions. Posttranslational modification of one or both subunits may occur by deamidination, resulting in multiple forms of the enzymes and creating a complex multibanded pattern on electrophoresis. 

The criterion for purity was a single band on a heavily loaded SDS-polyacrylamide electrophoresis gel. Antibodies were raised in rabbits in response to three intravenous injections each of 2mg. of laccase A with Freunds adjuvant, at 2 weekly intervals. Blood was removed 7d after the last injection and serum collected after coagulation of the blood cells. 

The second main class of blood constituents used as genetic markers are the polymorphic enzymes. The enzymes of interest to the forensic serologist are primarily located within the red blood cell and are commonly referred to as isoenzymes. These can briefly be described as those enzymatically active proteins which catalyze the same biochemical reactions and occur in the same species but differ in certain of their physicochemical properties. (This description does not exclude the tissue isoenzymes that occur within the same organism however, our consideration deals only with those of the red blood cell in particular.) The occurrence of multi-molecular forms of the same enzyme (isoenzymes) has been known for several decades however, it was not until the Metropolitan Police Laboratory of Scotland Yard adapted electrophoretic techniques to dried blood analysis that these systems were catapulted to the prominence they presently receive (.2). For many of the forensic serologists in the United States, the use of electrophoresis and isoenzyme determination is a recently-inherited capability shared by only a few laboratories. 

Markuszewski, M.J., Szczykowska, M., Siluk, D., Kaliszan, R. Human red blood cells targeted metabolome analysis of glycolysis cycle metabolites by capillary electrophoresis using an indirect photometric detection method. J. Pharm. Biomed. Anal. 39, 636-642 (2005)... 

Talamo F, D Ambrosio C, Arena S, Del Vecchio P, Ledda L, Zehender G, Ferrara L, Scaloni A. Proteins from bovine tissues and biological fluids Defining a reference electrophoresis map for liver, kidney, muscle, plasma and red blood cells. Proteomics 2003 3(4) 440-60. 

Theodorakis, C.W., D Sumey, S.J. and Shugart, L.R. (1994) Detection of genotoxic insult as DNA strand breaks in fish blood cells by agarose gel electrophoresis. Environ. Toxicol. Chem., 13, 1023-1031. 

Platsoucas and Catsimpoolas (1978) made the observation that peripheral rat blood cells can be separated by electrophoresis in 80% pure B... 

A.R. Minerick, R. Zhou, P. Takhistov and H.-C. Chang, Manipulation and characterization of red blood cells with alternating current fields in microdevices. Electrophoresis, 24(21), 3703-3717 (2003). 

H. Hwang, Y.-J. Choi, W. Choi, S.-H. Kim, J. Jang and J.-K. Park, Interactive manipulation of blood cells using a lens-integrated liquid crystal display based optoelectronic tweezers system. Electrophoresis, 29(6), 1203-1212 (2008). 

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