Capacitance measurements were conducted to determine the dielectric constant of electrostatic self assembly (ESA) films over various temperature and frequency ranges at 1 mV and 15% relative humidity. Measurement electrodes were fabricated on the ESA films using silver grease and a brass bar. In these tests, polymeric nanofilms were fabricated on gold-coated glass slides using the ESA process. Thicknesses of the films were between 100 nm and 600 nm obtained by depositing different number bilayers of negatively charged Poly s-119 (PS-119) or heparin and positively charged poly(diallyldimethylammonium chloride) (PDDA). The test results showed that dielectric constant values were around 2. Based on the test results, we concluded that this is a technique that might prove useful to estimate the capacitance and dielectric constant values of nanostructured ESA films, which can be largely used in the near future.