Background. Gangliosides act as important roles in tumor progression. B4GALNT1 is a key enzyme in ganglioside biosynthesis. B4GALNT1 expression is linked to tumorigenesis and the prognosis of tumor patients. Nevertheless, the role of B4GALNT1 in pan-cancer remains unclear. Methods. Several databases, including TCGA, GEO, GTEx, NCI-60, and TIMER, were searched. Methods including correlation analysis, Cox regression analysis, and Kaplan-Meier analysis were used to explore the expression pattern, prognosis, tumor infiltration pattern, genetics and epigenetics, and drug sensitivity of B4GALNT1 in pan-cancer patients from the above datasets. Results. B4GALNT1 was found to be aberrantly expressed in multiple types of tumors. The survival status of tumor patients was significantly related to B4GALNT1 expression, but the correlations were tumor-specific. Moreover, the expression of B4GALNT1 was associated with ImmuneScore and StromalScore in 21 and 27 tumor types, respectively. Also, B4GALNT1 was significantly associated with TMB, MSI, MMR, and DNA methylation. Additionally, the sensitivity of 9 drugs was correlated with the expression of B4GALNT1. Conclusion. A correlation of B4GALNT1 expression with prognosis exists in multiple types of cancers. In addition, B4GALNT1 expression may play a role in TME and tumor immunity regulation. Further investigation of the biological mechanisms of its different roles in tumorigenesis and clinical application as a biomarker is still required.