Like the other axioms of countability, separability is a "limitation on size", not necessarily in terms of cardinality (though, in the presence of the Hausdorff axiom, this does turn out to be the case; see below) but in a more subtle topological sense. In particular, every continuous function on a separable space whose image is a subset of a Hausdorff space is determined by its values on the countable dense subset.
A simple example of a space that is not separable is a discrete space of uncountable cardinality.
The property of separability does not in and of itself give any limitations on the cardinality of a topological space: any set endowed with the trivial topology is separable, as well as second countable, quasi-compact, and connected. The "trouble" with the trivial topology is its poor separation properties: its Kolmogorov quotient is the one-point space.
Separability is especially important in numerical analysis and constructive mathematics, since many theorems that can be proved for nonseparable spaces have constructive proofs only for separable spaces. Such constructive proofs can be turned into algorithms for use in numerical analysis, and they are the only sorts of proofs acceptable in constructive analysis. A famous example of a theorem of this sort is the Hahn–Banach theorem.