The Ly-3 antigens on mouse thymocytes: immune precipitation and molecular weight characterization

Specific anti-Ly sera were employed to precipitate Ly antigens from Nonidet P-40 extracts of mouse thymocytes labeled with 125I using lactoperoxidase and with NaB3H4 using galactose oxidase. Thymocytes from mice of the congenic strains C57BL/6J (Ly-2.2, Ly-3.2 positive), C57BL/6Ly-2a, Ly-3a (Ly-2.1, Ly-3.1 positive) and C57BL/6-Ly-2a (Ly-2.1, Ly-3.2-positive) were used as sources of labeled antigens and as immune adsorbants to permit evaluation of the specificity of each anti-Ly serum employed. Results of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) under reducing conditions are consistent with the Ly-3.1 antigen containing a glycoprotein subunit with an apparent mol wt of 35,000 daltons. Specific precipitates obtained using anti-Ly-2.1 serum yielded SDS-PAGE profiles identical to that obtained with anti-Ly-3.1 serum, suggesting that the Ly-2 and Ly-3 antigens have the same molecular weight distribution. The relationships of these results to the observed close genetic and topological linkage of Ly-2 and Ly-3 and to the genetic linkage of these loci with the IB-peptide marker, a mouse Bk-region polymorphism, are discussed.