Two-dimensional immunoelectrophoresis

Vl. Versey, J., An automated system of two-dimensional Immunoelectrophoresis. Protides Biol. Fluids, Proc. Colloq. 19, in press (1971). 

For the purpose of two-dimensional immunoelectrophoresis agarose slab gels are transferred from one plate to another one by using long razor blades. If narrow slits are needed, slit formers are used. The slit formers are positioned in such a way that... 

Scherer, R., M. Abd-El-Fattah, and G. Ruthenstroth-Bauer. 1977. Some applications of quantitative two-dimensional immunoelectrophoresis in the study of the systemic acute-phase reaction of the rat. In Perspectives in inflammation, future trends and developments, ed. 

A number of developments to further improve the resolution and specificity of the method have been reported [78,79], namely cross-over electrophoresis, rocket electrophoresis, two-dimensional Immunoelectrophoresis, and radioimmunoassay. 

C8. Converse, C., and Papermaster, D. S., Membrane protein analysis by two-dimensional immunoelectrophoresis. Science 189, 469-471 (1975). 

The nomenclature 2D6EL" generally has come to mean a combination of lEF and SDS-PA6E as described above. An example of such a separation of renal cell carcinoma Is shown in figure 1. Over 1000 spots are visible on this silver-stained gel. While this discussion will be limited to this particular technique, the reader should be aware that many two-dimensional electrophoretic techniques have been utilized, such as two-dimensional Immunoelectrophoresis and two-dimensional SDS-PA6E ( e.g.,+/- 2- mercaptoethanol). 

The strict differences observed in the antigenic spectrum of proteins in wild-type and mutant lethal giant larval of D. melanogaster do not correlate well with a relatively good similarity observed in the puffing activities of these strains. The protein differences were observed with the aid of two-dimensional Immunoelectrophoresis (Karakin, personal communication). 

P2b. Peeters, H., and Vuylsteke, P., Immunoelectrophoresis of two-dimensional patterns. Protides of the Biological Fluids, Proceedings of the Seventh Colloquium. Elsevier, Amsterdam, 1959. In press. 

Activation of C3 has been determined in the past by either immunofixation or two-dimensional, or crossed, Immunoelectrophoresis of plasma, with determination of the relative amounts of C3 and C3c. Currently, assays of fragments using antisera to neoaintigens (such as those on C3a) arfe preferred for this determination. For either type of assay, plasma samples must be obtained with precautions to prevent in vitro activation by plasmin, Cls, and leukocyte proteases The first few milliliters are discarded (or used for other purposes), then whole blood is collected through the same needle into a tube containing EDTA, Centrifugation should be performed as soon as possible and the separated plasma frozen below -40 C. 

Immunoelectrophoresis [123] can be characterized as two-dimensional agarose gel electrophoresis in which, in the second dimension, the advantage of immunoprecipi-tation reactions are utilized. There are five different arrangements of this technique in use today [124-127] (1) crossed Immunoelectrophoresis (2) fused rocket im-munoelectrophoresis (3) rocket immunoelectrophoresis (4) cross line immunoe-lectrophoresis (5) tandem crossed immunoelectrophoresis. For more detailed reviews see Refs. 128, 129. In all variations the important aspect is the possibility of obtaining quantitative data. 

Gel procedures originate from the classical immunoelectrophoresis of P. Grabar, often referred to as lEA, an abbreviation for immunoelectrophoretic analysis, and from the crossing diagrams of K. Nakamura. A distinction may be made between the two-dimensional QE of Clarke and Freeman (Figure 1) and the one-dimensional rocket immunoelectrophoresis of Laurell, often referred to as electroimmunoassay (EIA). Numerous technical modifications of these methods have been devised in order to obtain a certain characterization of one or more proteins. The methods are suitable for establishing an inventory of proteins and for molecular characterization of... 

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