Polyclonal and monoclonal antibodies were raised against a synthetic peptide containing the 15 carboxy‐terminal amino acids (497‐511) of vesicular stomatitis virus glycoprotein (VSV‐G). The polyclonal antibodies (alpha P4) reacted with epitopes distributed along the 15‐residue peptide, whereas the monoclonal antibody (P5D4) reacted with one epitope containing the five carboxy‐terminal amino acids. Both types of antibodies recognized the cytoplasmic domain of VSV‐G synthesized by tissue culture cells infected with the temperature‐sensitive 045‐VSV mutant (ts045‐VSV). They recognized immature forms of VSV‐G in the rough endoplasmic reticulum (RER) and Golgi complex, as well as mature VSV‐G at the cell surface and in budding virus. The effect of these antibodies on intracellular transport and maturation of VSV‐G was studied by microinjection. Both divalent antibodies (alpha P4 and P5D4) blocked transport of VSV‐G to the cell surface. Monovalent Fab' fragments of alpha P4 (alpha P4‐Fabs) also interfered with the appearance of VSV‐G at the cell surface; Fab fragments of P5D4 (P5D4‐Fabs), however, had no inhibitory effect. These results suggest that accessibility of a cytoplasmic domain, located within the sequence of amino acids 497‐506 of the carboxy‐terminal tail, is essential for transport of VSV‐G to the cell surface.