A new Ag(I) adduct was synthesized by the reaction of 4-amino-4H-1,2,4-triazole (L) with AgNO3. Its chemical structure was approved to be [Ag2(L)2(NO3)]n(NO3)n utilizing elemental analysis, FTIR spectra, and single crystal X-ray diffraction (SC-XRD). According to SC-XRD, there are two independent silver atoms which are coordinated differently depending on whether the nitrate anion is coordinated or not. The coordination geometry of Ag1 is a slightly bent configuration while Ag2 has a distorted tetrahedral structure. The 4-amino-4H-1,2,4-triazole ligand and one of the nitrate groups adopt bridging mode, which connects the crystallographically independent Ag1 and Ag2 atoms resulting in the formation of two-dimensional coordination polymer. Hirshfeld surface analysis displays that the intermolecular O···H (34.0%), Ag···N (10.6%), H···H (10.4%), Ag···O (9.3%), and N···H (9.0%) contacts are the most abundant interactions. Regarding anticancer activity, the [Ag2(L)2(NO3)]n(NO3)n demonstrates stronger cytotoxic efficacy against lung (IC50 = 3.50 ± 0.37 µg/mL) and breast (IC50 = 2.98 ± 0.26 µg/mL) carcinoma cell lines than the anticancer medication cis-platin. The [Ag2(L)2(NO3)]n(NO3)n complex showed interesting antibacterial and antifungal activities compared to the free components (AgNO3 and 4-amino-4H-1,2,4-triazole). The investigated silver(I) complex exhibits remarkable antibacterial activity against E. coli (MIC = 6.1 µg/mL) that may be on par with Gentamycin (MIC = 4.8 µg/mL). As a result, the newly synthesized Ag(I) complex could be suggested for anticancer and antibacterial treatments.