Rocket electrophoresis

Figure 7.6 Rocket electrophoresis. At pH 8.6 most proteins move towards the anode and precipitation occurs where the antigen and the antibody (in the gel) are in equivalent proportions. The size of the resulting rocket -shaped pattern of precipitate is proportional to the original concentration of antigen. The immunoglobulins show very little electrophoretic mobility at pH 8.6 and so remain in the gel during the process.

Electroimmunoassay (rocket electrophoresis) and radial immunodiffusion (A5) lack sensitivity at low Lp(a) concentrations, and the response is influenced by the size of the apo(a) isoforms (A5, K28). Differences in migration velocity in the agarose gel lead to an underestimation of the samples with large apo(a) isoforms and to an overestimation of samples with small apo(a) isoforms. Moreover, the detection limit lies around 0.07-0.08 g/liter Lp(a), so that this method is better suited for screening and detection of individuals with elevated Lp(a) levels than for the exact measurement of the plasma Lp(a) concentration. 

Receptor mimics (anti-idiotypic antibodies) Immunoelectrophoresis (rocket electrophoresis)... 

Radial immunodiffusion and double diffusion experiments can be accelerated by electrophoresis. In rocket electrophoresis, so-called because of the shape of the zone of precipitation, electrophoresis of proteins is carried out in an agarose gel which contains antibody. The precipitated antibody-antigen complexes form rocketshaped zones when the pH is adjusted (often pH 8.0) so that antigen and antibody migrate in opposite directions (Figure 6-6)... 

Figure 10.14. Principles of rocket electrophoresis. Rocket height is measured from the cathodic end of the well to the rocket tip, and is directly proportional to antigen concentration.

Many modifications have been made since the original technique was described, several of them permit quantitative evaluation. The most common of these methods are rocket electrophoresis, crossed electrophoresis, jused rocket electrophoresis, intermediate gel electrophoresis, and tandem electrophoresis. The basic arrangement for each is described below. 

Suppose you have collected several fractions from a gel filtration, ion exchange, chromatographic, or any other similar separation and want to know in which fraction/s the compounds of interest are. Figure 28-8 shows the spotting template and the results. Because rockets from several fractions may diffuse together, this is known bs fused rocket electrophoresis. The black dotted sample wells in Figure 28-8 are fractions that have been pooled before the run. 

Figure 28-8. Template for fused rocket electrophoresis and a set of results.

Two major variations of the method are used at present. The first approach is closely related to some immunoelectrophoretic techniques, e.g., crossed Immunoelectrophoresis or rocket electrophoresis [180,181], the only difference is that usually a lectin instead of an antibody is incorporated in the agarose gel the conditions in the gel (electroendoosmosis and pH) are used such that the lectin has a very low electrophoretic mobility. The lectin reacts with specific glycoproteins and forms characteristic precipitation lines analogously to the reactions observed during Immunoelectrophoresis between the antibody and the antigen. This method can be... 

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. 

Fig. 3. Immunological reactions, where Ag is antigen and Ab is antibody, for detection in electrophoresis (a) Ouchtedony technique (b) single-radial diffusion (c) rocket immunoelectrophoresis and (d) crossed immunoelectrophoresis.

Use (Solid) Mechanical rubber products, lining oilloading hose and reaction equipment, adhesive cement, binder for rocket fuels, coatings for electric wiring, gaskets and seals. (Liquid) Specialty items made by dipping or electrophoresis from the latex. (Foam) Adhesive tape to replace metal fasteners for automotive accessories, seat cushions, carpet backing, sealant. 

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. 

Relevant for the lateral electron transport is mobile plastotyanin. Reduced plastocyanin bound to PS I reduces oxidized P-700 after a short flash with a characteristic halftime of 10-14 ns [17]. A measurement (not shown) in a dark-adapted sample indicates that to 95 % of total PS I a plastocyanin molecule is bound. We have determined a molar ratio of 205 for chlorophyll to plastotyanin by rocket immuno-electrophoresis and from the amplitude of the P-700 signal a molar ratio of 3.3 for plastocyanin to P-700. These data and the values in Table 1 suggest that in the dark... 

For this reason we developed a specific quantitative micromethod, the rocket analysis or immunoelectrodiffusion, for both CF and S, without the need for previous extraction and independent of purity, activity, and several chemicals usually present in biochemical samples (Berzborn et al. 1981 Roos,Berzborn, 1983). For electroimmunodiffusion with both our antisera (s.below) peptide S had to be dissociated from CF by deoxycholate before the electrophoresis (Roos,Berzborn, inprep.). 

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