Anti-Thy-1 PFC assay

NY) and kept at 4°C in the dark until developed. The time of exposure varied between 12 days and one month. The films were developed in Kodak X-ray Developer-Replenisher ( 146-5327) and fixed in Kodak Rapid Fix ( 146-4106). Selected areas on the TLC plates directly under the spots on the films were eluted from the silica gel and tested in the anti-Thy-1 PFC assay. 

Anti-Thy-1 PFC Assay. Fuji and Milgrom (4) originally developed an in vitvo PFC assay which detected Thy-1 alloantigen on whole thymocytes. A modified version, used here, has previously been described in detail and found to be effective for measuring the immune response to isolated glycolipid and glycoprotein Thy-1 alloantigens (3). 

Chromatography was done as described in Figure 3, using the gangliosides derived from 2 X 10s cells. Assayed fractions are labeled with numbers that correspond to the anti-Thy-1 PFC assay in Figure 6. Number 10 refers to the area surrounding the spots that were assayed (15,). 

Thy-1 glycolipids isolated from the two-dimensional TLC plates in Figures 3 and 5 were tested for allogenic specificity with the anti-Thy-1 PFC assays. Values are the mean standard error of five cultures. Application of the Student t test to the standard errors for these samples gives p values less than 0.05 when compared wtih the Thy-1-active fraction (15/... 

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