Background: The aim of this study was to discover unique membrane proteins associated with gastric cancer (GC) in proteomics analysis. Methods: Using a data-independent acquisition strategy, we compared the relative expression levels of membrane proteins in GC. Results: A total of 2774 differentially expressed membrane proteins were identified between GC and normal cell lines. Conjoint analysis of transcriptomes and proteomes provided 11 potential biomarkers (GPRC5A, PSAT1, NUDCD1, RCC2, IPO4, FAM91A1, KANK2, PRADC1, NME4, METTL7A and LRP1B) for further exploration. Downregulation of LRP1B in GC was validated by immunohistochemistry. Moreover, LRP1B demonstrated an area under the receiver operating characteristic curve of 0.917 in differentiating GC from normal tissues. Conclusion: LRP1B was identified as a meaningful indicator assisting in GC detection and labeling of tumor boundaries.