In molecular charge transport, transition voltage spectroscopy (TVS) holds the promise that molecular energy levels can be explored at bias voltages lower than required for resonant tunneling. We investigate the theoretical basis of this tool using a generic model. In particular, we study the length dependence of the conducting frontier orbital and of the “transition voltage” as a function of length. We show that this dependence is influenced by the amount of screening of the electrons in the molecule, which determines the voltage drop at the contacts or across the entire molecule. We observe that the transition voltage depends significantly on the length, but the ratio between the transition voltage and the conducting frontier orbital is approximately constant only in strongly screening (conjugated) molecules. Uncertainty about the screening within a molecule thus limits the predictive power of TVS. We furthermore argue that the relative length independence of the transition voltage for nonconjugated chains is due to strong localization of the frontier orbitals on the end groups ensuring binding of the rods to the metallic contacts. Finally, we investigate the characteristics of TVS in asymmetric molecular junctions. If a single level dominates the transport properties, TVS can provide a good estimate for both the level position and the degree of junction asymmetry. If more levels are involved, the applicability of TVS becomes limited.