Agar gel electrophoresis

H8. Hirschfeld, J., A simple method of determining haptoglobin groups in human sera by means of agar-gel electrophoresis. Acta Pathol. Microbiol. Scand. AT, 169 (1959). 

A different procedure to avoid pH changes and also loss of potential over the baffles is to add fresh buffer continuously to the electrode vessel, which has an overflow. Thus the electrodes lie far nearer the paper without an interposed baffle system. Grabar introduced this method for agar gel electrophoresis (G9). We ourselves use a pump to circulate the buffer continuously over the two electrodes, catching the overflow in one common reservoir, where it mixes and then returns to circulation. This system proved particularly useful in continuous two-dimensional electrophoresis, as will be seen in Part II. But it will also help in the building of easily standardized electrophoretic chambers, necessary for standardization of clinical work on electrophoresis of proteins. 

The recent fractionation work of Seijffers et al. (S17-S19b) on DEAE-cellulose and that of Grabar and Burtin s laboratory by Rapp et al. (Rl), Kushner et al. (K32), and Hirsch-Marie et al. (H20b) utilizing agar gel electrophoresis and immunoelectrophoresis, resulted in differentiation of 3 to 4 proteases in human gastric juice, each having its own proenzyme. Detailed discussion of this work is presented in the companion review (G21) of the author, in this volume. 

Rapp with Burtin (R1) found 5 carboxylesterases in the gastric human mucosa on agar gel electrophoresis (see Fig. 11) using special substrates. These carboxyl esterases were decreased and differently distributed in the gastric mucosa of patients with gastric cancer. 

Fig. 19. Agar gel electrophoresis of proteins of concentrated gastric juice after in vivo alkalinization. Top row Normal human serum. Bottom row Concentrated gastric juice. From Hirsch-Marie and Burtin (H7).

Hirsch-Marie and Burtin (H7a) determined proteolytic activity of human gastric juice after histamine by electrophoresis on agar, using, as a substrate, human or bovine serum albumin at 0.2% concentration in a glycine-HGl buffer of 0.2 molarity and pH 2. Acid gastric juices with pH below 5.8 showed 4 proteolytic constituents on agar gel electrophoresis. These corresponded in their pH optimum of activity and in immuno-... 

Slow anodic binders of Gras-beck—on starch block electrophoresis, GuUberg—on paper electrophoresis, Jeffries et al.—on starch gel electrophoresis, Okuda and Grasbeck, and Simons and Grasbeck—on agar gel electrophoresis (binder S). 

Fast anodic binders of Grasbeck on starch block electrophoresis, and Jeffries et al. on starch gel electrophoresis as well as intermediate binder of Simons and Grasbeck on agar gel electrophoresis (binder I). 

Fast anodic binder of GuUberg on paper electrophoresis of gastric juice neutralized in situ, and rapid anodic binder of Okuda and Grasbeck, and Simons and Grasbeck on agar gel electrophoresis (binder R). 

Menard L, Dempsey ME, Blankstein LA, Aleyassine H, Wacks M, Soeldner JS. Quantitative determination of glycosylated hemoglobin A1 by agar gel electrophoresis. Clin Chem 1980 26 1598-602. 

Significant advances have been made during the last decade in the development of new techniques of paper, starch-, and agar-gel electrophoresis. Soon after the introduction by Smithies in 1955 of the technique of starch-gel electrophoresis as an important analytical tool for the fractionation of serum proteins (P21, R8), this procedure proved val-... 

Stevenson (S47) and Port and Van Venrooy (P14) were able to demonstrate alkaline phosphatase activity following agar-gel electrophoresis. This technique was further developed by Haije and DeJong (HI), who obtained characteristic separate bands for liver and bone alkaline phosphatases. Dymling (D24) applied the technique to pregnancy serum and placenta. 

HI. Haije, W. H., and DeJong, M., Iso-enzyme patterns of serum alkaline phosphatase in agar-gel electrophoresis and their clinical significance. Clin. Chim. Acta 8, 620-623 (1963). 

Stevenson, D. E., Demonstration of alkaline phosphatase activity following agar-gel electrophoresis. Clin. Chim. Acta 6, 142-143 (1961). 

W25. Wieme, R. J., An integrated procedure for agar gel electrophoresis. Protides Biol. Fluids, Proc. Colloq. 11, 398-400 (1964). 

D2, Dahl-Jorgensen, K., and Larsen, A. E., Hb Aj determination by agar gel electrophoresis after elimination of labile Hb Aj a comparison with ion-exchange chromatography, Scand. ]. Clin. I b. Invest. 42, 27-33 (1982). 

Comprehensive books on agar gel electrophoresis have been written by Wieme (W3, W4). For detailed information on the subject the reader is referred to diese books. Agar gel is mostly used as a 1% solution in a buffer solution. For ultramicro electrophoresis it is used in a layer of 1 mm thickness, which may be mounted on a microscope slide. 

W3. Wleme, R. J., Studies on agar gel electrophoresis. Tedmiques-applications. Thesis Ghent, Arsica Uitgaven N. V., Brussels, 1959. 

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