Crossed line electrophoresis

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. 

Fig. 3. Crosslinking of l25I-labelled human growth hormone to lactogenic receptors from rat liver and the Nb2 cell line. Hormone-receptor complexes were cross-linked with disuccinimidyl suberate, fractionated by SDS gel electrophoresis and detected by autoradiography. Lanes 1-4, Nb2 cells lanes 5-8, rat liver. Lines on the left indicate the positions of the hormone-receptor complex for Nb2 cells (102 kDa), rat liver (71 kDa) and uncomplexed hormone (hGH). Numbers on the right are M, values for standards (kDa). Samples were incubated with (lanes 2,4,6,8) or without (lanes 1,3,5,7) unlabelled hGH and electrophoresed under reducing (lanes 1,2,5,6) or non-reducing (lanes 3,4,7,8) conditions. From Ref. 36.

Instrumentation. A Rank Bros, micro-electrophoresis unit was used in those studies, with a specially made quartz cell having a 6 cm. path length of rectangular inside cross-section (l mm thick, 10 mm deep) in which the Komagata equation (25) predicts zero mobility of the liquid phase in planes located at 0.612 of the distance b from the center plane of the cell to the wall. In electrophoresis experiments 300 to 1200 volts were applied to the cell and mobilities measured in planes a distance h from the center plane. The results were graphed as observed velocity versus (h/b)z as proposed by van Gils (26J, and if the straight lines characteristic of perfect parabolic flow resulted, the electrophoretic mobilities (v ) observed at h/b=0.612 were considered acceptable for calculation of zeta-potential. Zeta-potentials were calculated by the Huckel equation (27) ... 

At about fhe same time, lin and coworkers [84] at fhe National Cheng Kung University in Tainan, Taiwan published fheir version of a chip, which is similar to that of Harrison s group in Alberta. This group had extensive experience in flow injection analysis and on-line analysis wifh conventional electrophoresis. In their design, a 3 mm wide X 40 pm deep sample inlet channel (SIC) was etched for sample flow-through, which in turn was connected to fhe electrophoretic manifold of 80 X 40 pm cross section, in a configuration similar to the Alberta chip (see... 

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... 

Fig. 1 Illustration of flexible polymers in dilute solution ic < c ), entanglement threshold (c = c ), and semidilute solution (c > c ) regimes, as well as cross-linked chain. One chain is drawn as a thick line for easier visualization. The small circle in the semidilute solution regime represents the blobs of size 4-Source From The separation matrix, in Analysis of Nucleic Acids by Capillary Electrophoresis ...

In a separate study. Rill and Al-Sayah examined two-dimensional electrophoresis of myoglobin tryptic polypeptides(47). The first dimension used a cross-linked polyacrylamide gel as the support medium. Advantage was then taken of the liquid nature of the cold triblock copolymer solutions to eliminate interface issues the cross-linked gel was simply pushed into the triblock system. With few exceptions, after the second dimension of electrophoresis, most peptides lie on a diagonal line, indicating that the predominant separation mechanism is the same in both media Al). 

Fig. 16. Immuno-electrophoFetic separation of a normal human serum and sketch of the portions of resulting precipitation lines. The cross-hatched circle indicates the point of application for electrophoresis. Antibodies were applied in a trough-shaped indentation at the lower border.

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