The present study was carried out to evaluate the potential antimicrobial activity of latex serum extracts (ethanol, methanol, chloroform and aqueous) of Calotropis Procera, against some reference pathogenic strains of bacteria and fungi, Results revealed that the latex extracts showed a vary ing degree of inhibition against all tested microorganisms. The aqueous and ethanolic extracts were more effective antimicrobials against all tested strains, while chloroform extract showed only antibacterial activity. In addition, Phytochemical analysis w ere also carried out to assess the presence of bioactive compounds in the plant latex serum. The analysis revealed the presence of Cardiac glycosides, Saponins, Phenolic compound, Terpenoids, Alkaloids, Flavonoids, Tannins, and Resins in some of the extrac ts. The concentration of the various classes of secondary metabolites varies amongst the extracts evaluated. The ethanolic and the aqueous extract were rich in Saponins, Terpenoids and flavonoids than in methanolic extract, while the chloroform extract was poor in their phytochemical contents where it contains only Cardiac glycosides and Terpenoids. The present investigation was concerned also with the synthesis of latex silver nanoparticles (LAg -NPs) using the latex of Calotropis procera as reducing and ca pping agent, the synthesized LAg -NPs were characterized using UV -VS spectrophotometer analysis, Fourier Transform Infrared Spectroscopy (FTIR), X-ray diffraction (XRD) and Transmission Electron Microscopy (TEM). Further, the antimicrobial effect of the syn thesized silver nanoparticles was determined against the reference pathogenic strains and compared with the effect of crude latex and latex solvent by agar well diffusion methods. The synthesized nanoparticles exhibited a variable growth inhibition against the tested strains. A significant increase in the effect of nanoparticles compared with crude latex and latex serum.