A new methodology has been developed to measure the maximum pressure that can be withstood by a bilayer lipid membrane (BLM) formed over porous substrates. A custom test fixture was fabricated to pressurize BLMs in very fine increasing increments until they fail. This experiment was performed on 1-Stearoyl-2-Oleoyl-sn-Glycero-3-Phosphatidylocholine (SOPC) BLMs formed over polycarbonate substrates with a single pore ranging from 5 to 20 microns in diameter. Failure pressure was found to be inversely proportional to pore diameter. The same set of experiments was repeated for BLMs that were formed from a mixture of SOPC and 50 mol% cholesterol (CHOL). The presence of cholesterol was found to increase the failure pressure of the BLMs by 56% on average. A model of the characteristic pressure curve from this experiment was developed based on the pressurization and flow of fluid through a porous substrate. The model was found to accurately fit the experimental pressure curves.