On Malignolipin. XII

Abstract

Comm. by Y. KUNO, M.J.A.., Oct. 12, 1961) In the preceding report, ' it was demonstrated that the subcutaneous injection of malignolipin resulted in the production of a complement combining specific antibody, which reacts only with malignolipin.To ascertain further the specificity of this antibody, and also to find an immunological method for the histological detection of malignolipin, some properties of fluorescent anti-malignolipinserum were examined.The anti-malignolipin rat and rabbit sera, which were prepared in the way described in the preceding report' and which were ascertained to have a complement-fixing titer of X 320, were treated with fluorescein isocyanate to make fluorescent sera according to Coons' method.2'The fluorescent sera were then absorbed with acetone-dried powder of normal liver of mice or rabbits to remove non-specific staining of normal tissues.2'The sera were freeze-dried for storage, and before using were diluted to X 5-10 with a buffered saline solution (0.15 M NaCI containing 0.01 M phosphate, pH 7.0). The complement fixing titer of the sera suffered almost no change by these procedures.An acetone-fixed frozen section of tissues was kept in a buffered saline solution for one hour, and stained on a glass slide with a drop of fluorescent sera for one hour at 37°C, care being taken to prevent evaporation.Then it was washed gently but thoroughly with a buffered saline solution, mounted with a drop of glycerol containing one part in 10 of buffered saline, and observed under a fluorescence microscope.Smears of blood, of ascites, and of serum or egg white were dried by air for 15 min at room temperature, fixed in acetone for 3 min, and at once stained with a drop of fluorescent sera for 10