Peptide arrays are a widely used tool for drug development. For peptide-based drug design it is necessary to screen a large number of peptides. However, there are often difficulties with this approach. Most common peptide synthesis techniques are able to simultaneously synthesize only up to a few hundred single peptides. Spot synthesis is a positionally addressable, multiple synthesis technique offering the possibility of synthesizing and screening up to 10,000 peptides or peptide mixtures on cellulose or other membrane surfaces. In this chapter we present the basic procedures and screening methods related to spot synthesis and outline protocols for easy-to-use detection methods on these peptide arrays.