Anti-lactose antibodies agar diffusion

Fig. (9). Inhibition of anti-lactose antibodies by increasing amounts of methyl P-lactoside and lactose (A) and by the agar diffusion method (B).

Fig. (10). Agar diffusion of different proteins and a lipid conjugated with lactose against anti-lactose antibodies (well L) and against anti-BSA antibodies (Well BS) wells 1-6 contained Lac-poly, Lac-BSA, Lac-sphingosine, Lac-ORA, Lac-HGG and BSA Chemical modification of the antigen by periodate oxidation or borohydride reduction can effect an agar diffusion against anti-gum arabic antibodies (Se), GA=gum arabic Bl=blank.

Fig. (15). Agar diffusion (AX electrophoresis (B) and isoelectric focusing (C-F) of anti-lactose antibodies.

Fig. (16)l Liquid dectrofocused pattern (A) electrophorised in gels stained by protein reagent (B) and agar diffusion (C) of anti-lactose antibodies. 

Sepharose and elution with lactose (Fig. 23). The UV-absorbing fraction that was eluted with the lactose solution was collected and the protein precipitated by addition of an equal volume of saturated ammonium sulfate. On refrigeration of the sample overnight, a white precipitate formed. This precipitate was collected by centrifugation and redissolved in 0.2 mL of 0.02 M phosphate buffer of pH 7 in saline. The agar-diffusion test shown in the inset of Fig. 23 showed that high-titer antibodies were produced that formed a precipitin complex with the antigen. Several affinity runs were made. The antibody samples were combined and lyophilized. The yield of anti-lactose antibody was 50 mg from 20 mL of serum. 

Fig. 25.—Gel electrophoresis and isoelectrofocusing of anti-lactose antibodies followed by agar diffusion. A, electrophoresis gel 1 stained with Coomassie Blue, gel 2 embedded in agar, area of precipitin formation (3), and solution of antigen (4). B, electrofocusing gel patterns (5-8) similar to series (1-4).

That both components were antibodies was established by agar diffusion tests with these preparations and solutions of the glycan performed in the usual manner. The results of the agar diffusion tests are shown in Figure 3, right plate. The two preparations of antibodies have been designated as anti-galactose (anti-gal) antibodies and anti-lactose (anti-lac) antibodies and some of the properties of the two sets of antibodies are described in a later section. 

The specificity of the antibodies can be verified by inhibition tests using agar diffusion data [35], The results of such experiments with anti-lactose... 

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