Immunoglobulin heavy chain locus variable gene segment V4-34 (V(H)4.21) use in productive heavy chain (IgH) gene rearrangements has been described in a number of human reactive and autoimmune B cell responses, and has been shown to be frequently used in some series of cases of diffuse large cell lymphoma (DLCL). The 9G4 antibody is relatively specific for the V4-34 gene product and can be used to screen for cells that use V4-34 in their productive IgH locus rearrangements. The purposes of this study were to determine the sensitivity of the 9G4 antibody against DLCL cases known to use V4-34, then to screen a variety of human lymphoma types for 9G4 reactivity. Frozen tissue sections were cut from 118 cases of various human lymphomas. Generalized 9G4 membrane reactivity was identified in 78% of DLCL cases known to use V434. 9G4 reactivity varied by lymphoma type for the unknown cases, with diffuse large cell lymphoma (30%) and mantle cell lymphoma (28%) showing statistically significant differences (P < .001) from the expected value of 6% V4-34 positivity in peripheral blood B cells. This nonrandom increased utilization of V4-34 in productive IgH locus rearrangements supports the hypothesis that Ig binding specificity may play a role in lymphomagenesis.