In this paper, macroporous antimicrobial polymeric gels (MAPGs) functionalized with active quaternary ammonium cations attached to varying hydrocarbon chain lengths have been fabricated. Apart from the change in the alkyl chain length attached to the quaternary ammonium cation, the amount of crosslinker was also varied during the fabrication of the macroporous gels. The prepared gels were characterized using Fourier transform infrared spectroscopy, X-ray photoelectron spectroscopy, field emission scanning electron microscopy (FE-SEM) and swelling studies. In addition, the mechanical properties of the fabricated macroporous gels were studied using compression and tensile testing. The antimicrobial activity of the gels has been determined for Gram-negative bacteria (Escherichia coli, Pseudomonas aeruginosa) as well as Gram-positive bacteria (Bacillus subtilis, Staphylococcus aureus). Antimicrobial activity, as well as the mechanical properties of the macroporous gels, was found to be influenced by the alkyl chain length attached to the quaternary ammonium cations as well as by the amount of crosslinker used for the fabrication of the gel. In addition, on increasing the alkyl chain length from C4 (butyl) to C8 (octyl), the effectiveness of the polymeric gels increased. It was observed that the gels derived using a tertiary amine (NMe2) containing monomer showed relatively low antimicrobial activity as compared to the gels obtained using quaternized monomers (C4 (butyl), C6 (hexyl), and C8 (octyl)). The gels based on the quaternized C8 monomer displayed the highest antimicrobial activity and mechanical stability as compared to the gels based on the C4 and C6 monomers.