Diagnostic methods and probes

The invention features methods for detecting a target analyte in a sample. In these methods, a sample is contacted with a detector probe and a capture probe to form a detector probe-analyte-capture probe complex. The detector probe includes a new moiety, having a predetermined pI and containing a detectable label. The complex is isolated from detector probe that is not bound in the complex, the moiety is released from the complex, and the released moiety is concentrated, e.g., by isoelectric focusing or other methods. Detection of the moiety, e.g., at a position in a pH gradient corresponding to its pI, can be used as a measure of the presence and concentration of the analyte in the sample.