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This study concerns sign and signification from the viewpoint of the three main parts of Stoic philosophy, i.e. logic, physics, and ethics. Its objective is to identify topics belonging to Stoic logic, physics and ethics which concern sign and signification in order to evaluate whether a general notion of sign applicable to Stoic philosophy as a whole may be discovered or constructed. The study is based on a corpus of ancient evidence and testimonia, drawing on surviving Stoic sources supplemented by second-hand sources, especially the works by Diogenes Laertius and Sextus Empiricus. The evidentiary value of Aristotelian and Epicurean philosophy is also discussed. As principal results of the study, pertinent topics in Stoic physics viz. sign and signification are identified, e.g. the concept of relativity in Stoic ontology and the Stoic doctrine of divination. In the context of Stoic logic, important distinctions related by Sextus Empiricus are analysed: an account of the sign as an essential concept in human rationality, and a threefold division consisting of the signifier, the signified and the existent. Sextus’ account concerning commemorative and indicative signification is then discussed, with evidence supporting the plausible attributability of these two kinds of signs to the Stoics being found in the resemblance between Sextus’ account of commemorative and indicative signification and Cicero’s testimony on Stoic divination. An outline of the general conception of sign is then presented through two properties, relativity and evidentiality. In a case study conducted in Stoic ethics, questions of moral psychology concerning knowledge of one’s ethical status are identified as of sign-inferences and conform to the general conception of sign. In conclusion, while several topics pertinent to sign and signification in Stoic philosophy yield, on closer analysis, support for a general conception of sign, construction of a more substantial account requires more research and a larger evidence-base. Prospects for further study in this regard are identified especially in the Stoic concept of pneuma.