Sixteen mouse and rat monoclonal antibodies reactive with gag or env proteins of Friend murine leukemia virus (F-MuLV) or recombinant MCF viruses related to F-MuLV were derived. Specificity of these was determined by immunofluorescence, immunoprecipitation, and reactivity with viral proteins blotted onto nitrocellulose paper. Seven antibodies reacted with envelope protein antigens of certain nonecotropic viruses only. Nine antibodies reacted with both ecotropic and nonecotropic viruses. Of this latter group, three were antienvelope, four were anti-p15, one was anti-p12, and one was anti-p30 in specificity. When tested as a panel against 10 strains of F-MuLV, these antibodies could distinguish seven different antigenic patterns. However, all 10 strains retained reactivity for three anti-gp70 antibodies uniquely specific for Friend and Rauscher MuLVs. Our antibody panel could also identify MCF viruses isolated from mice neonatally inoculated with F-MuLV as recombinants related to a particular F-MuLV strain based on identity of p15 gag antigenic profiles. However, recombinant viruses lacked several envelope antigens always associated with F-MuLV and instead had new envelope reactivities. These anti-MCF monoclonal antibodies detected no shared envelope antigens between MCF and xenotropic viruses isolated from mice inoculated with F-MuLV, however many of them did react with MCF viruses derived from AKR mice and NFS mice congenic for endogenous ecotropic virus loci.