Pure venom from 3 Indian spiders was extracted by electrical stimulation. The average amount of venom yielded per spider and the average protein content in the venom were evaluated. The venom proteins and free amino acids have been separated by electrophoretic and chromatographic techniques respectively. Nine major fractions were separated from the wolf spider,Lycosa indagastrix and 7 from funnel web spiderPleisiophrictus collinus and the common house spiderHeteropoda venatoria. Each venom had a distinct electrophoretic pattern. Chromatographic studies revealed that proline was the only free amino acid common to all the 3 venoms.