Antibodies specifically recognizing integral membrane protein are essential tool for functional analysis, diagnosis, and therapeutics targeting membrane proteins. However, development of antibodies against membrane protein remains a big challenge, because mass production of membrane protein is difficult. Recently, we developed a highly efficient cell-free production method of proteoliposome antigen using cell-free protein synthesis method with liposome and dialysis cup. Here we introduce practical and efficient integrated procedures to produce large amount of proteoliposome antigen for anti-membrane protein antibody development.