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• W4313307860 abstract "Summary Several conditions of storage of antisera to be used in quantitative determinations of antibody were investigated to determine their effect on the optical density measurements in gel precipitin techniques. Antisera fleshly stored at 4°C showed decreased O.D. after 3 days, Antisera stored at 25°C showed an initial decrease in O.D. followed by an increase which appeared maximal at 16 days, and, finally, a decrease in O.D. Antisera incubated at 56°C showed an initial decrease in O.D. followed by an increase in O.D. The initial decrease in O.D. following storage of antisera at 4°C or 25°C or heating of antisera at 56°C paralleled the loss of hemolytic complement activity of the antisera, the presence or absence of which was determined by complement fixation. The factor causing the initial O.D. decrease could be removed from the antisera by absorption with an immune precipitate. Also, preliminary experiments, not described here, show that the factor or factors responsible for the initial high O.D. found with fresh antisera could be removed by treating such antisera with either EDTA or zymosan. The increase in O.D. during storage at 25°C or incubation at 56°C appears to be due to denaturation or alteration of serum components in such a manner that they interact with the antigen antibody precipitation process to produce denser precipitates. A statistical comparison of the results of analysis of nine antisera using both the classical precipitin technique and the agar gel precipitin techniques, by means of Student's t test, indicated no difference in the means at the 5% level, provided O.D. corrections of zero to 0.02 units were made in the gel precipitin measurements. The agar gel precipitin technique offers considerable savings in both time and material, since an analysis can be obtained from O.D. measurements at a single concentration of antiserum after only 2 days of incubation at 25°C. The data presented here indicate that for routine use of this technique, one should: a) inhibit the hemolytic complement of all antisera to obtain standard curves with zero intercepts, b) store such decomplemented antisera at subzero temperatures to prevent further changes which might influence O.D. values and c) keep the antiserum-agar mixture in the reaction bath at 56°C for the minimal time needed to charge triplicate reaction tubes to prevent abnormal O.D. increases, particularly those observable at higher antiserum concentrations." @default.
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• W4313307860 date "1969-06-01" @default.
• W4313307860 modified "2023-10-16" @default.
• W4313307860 title "Antigen-Antibody Reactions in Agar" @default.
• W4313307860 doi "https://doi.org/10.4049/jimmunol.102.6.1398" @default.
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