Four alkaline ribonucleases [EC 3.1.4.22] were purified 2,050- to 3,460-fold from bovine submaxillary gland by repeated CM-Sephadex C-25 chromatography and Sephadex G-50 gel filtration, with a total recovery of about 13%. These were designated as RNase BS1, BS2, BS3, and BS4, based on their order of elution from a CM-Sephadex C-25 column. The molecular weights of these enzymes were estimated by gel filtration to be 19,000, 17,500, 17,000, and 12,000, respectively. These enzymes are very similar to RNase A in that they are inhibited by heparin, show preferential hydrolysis of C5'-O-P linkages adjacent to a cytosine nucleotide rather than a uracil nucleotide, and in their antigenic properties. Spermine was found to stimulate the activities of these enzymes; the degree of stimulation was in the order RNase BS4 greater than BS3 greater than BS2 greater than BS1. The stimulation by spermine is due to the increased cleavage of C5'-O-P linkages adjacent to cytosine nucleotide. The reason for the differences in the degree of spermine stimulation of these enzymes is discussed.