Rat mAb have been raised to mouse liver-derived large granular lymphocytes (LGL). One of these mAb (4D11) binds specifically to mouse LGL and appears to recognize a non-allelic determinant on NK-active cell populations. The Ag recognized by 4D11 is expressed on LGL of all mouse strains tested, including C57BL/6 (B6), BALB/c, C3H/HeJ, and SJL/J; thus, we have provisionally called this Ag LGL-1. Analysis of various lymphoid and hemopoietic tissues has indicated that only normal tissues known to contain NK activity have 4D11+ cells. With B6 and B6 congenic strains, a positive correlation exists between the number of LGL in a sample and the percentage of 4D11 immunofluorescence-positive cells detected by flow cytometric analysis. Dual color immunofluorescence analyses indicate that some LGL-1+ cells are also stained for Ly-1 and Thy-1. A very small subset exists that is weakly positive for CD3 and LGL-1. However, virtually no cells are seen which co-express LGL-1 and Ly-2. LU activity against YAC-1 targets was increased 7- to 700-fold in LGL-1+ spleen cells obtained by cell sorting from several different strains of mice (B6, BALB/c, C3H/HeJ, SJL/J, and athymic/nude). Sorted, LGL-1- spleen cells contained little or no NK activity. Cells positively selected for LGL-1 also contained between 50 and 60% LGL by morphology. By using facilitated in vitro antibody plus C' treatments, the majority of NK activity can be depleted from both B6 spleen and liver-derived leukocyte populations enriched for NK cells. mAb 4D11 was also shown to precipitate a protein of approximately 87 kDa from the surface of enriched murine NK cells. This mAb should prove valuable for understanding the role of NK cells in the immune response.