WELLS, ARTHUR F. (Space-General Corp., El Monte, Calif.), CURTIS E. MILLER, AND MARVIN K. NADEL. Rapid fluorescein and protein assay method for fluorescentantibody conjugates. Appl. Microbiol. 14:271-275. 1966.-A method is presented for the rapid determination of the fluorescein content, protein content, and fluorescein-to-protein ratio for immune globulin conjugates with fluorescein isothio-271

A method is presented for the rapid determination of the fluorescein content, protein content, and fluorescein-to-protein ratio for immune globulin conjugates with fluorescein isothiocyanate as the fluor. This method is based on the absorbance of the fluorescent antibody at those wavelengths primarily associated with the fluorescein and γ-globulin fractions, and permits these materials to be determined by a single nondestructive analytical procedure. A small sample of the fluorescent antibody, in many cases 0.1 ml or less, is adequate for the above determinations. A nomograph is presented which allows simultaneous determination of the materials from the observed absorbance at the two wavelengths. The method is sufficiently accurate for most applications of the fluorescent-antibody techniques. Although this procedure has been developed primarily for fluorescent-antibody conjugates prepared from rabbit γ-globulin, it can be used directly for antibodies prepared from other animals provided the γ-globulin is relatively free from albumin.

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Treponema pallidum can only be cultured in living animal tissue, such as rabbit testes. However, the extract of these organisms from the testicular material leaves the T. pallidum contaminated with tissue debris. This paper describes the separation of T. pallidum from the debris by continuous-particle electrophoresis. The importance of equilibration time before electrophoresis is discussed. Treponema pallidum at present can be propagated only in living animal tissues. Large numbers of T. pailidum can be obtained from rabbit testes inoculated with a rabbit-adapted strain-the Nichols strain. Practical usage of such T. pallidum for test antigens, vaccine material, or antigen studies necessitates removal of the testicular debris as efficiently and thoroughly as possible. Previous investigators have sought to accomplish this by various techniques of centrifugation, including differential centrifugation (2), tartrate gradients (3), and cesium chloride gradients in zonal rotors (Thomas et al., personal communica

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