Many ion channels are localized in areas of the plasma membrane enriched in cholesterol and sphingolipids, known as “lipid rafts.” The problem of the interaction of ion channels with rafts is one of the least studied in modern biology and physiology. In this study, we investigated the role of lipid rafts in the membrane localization of TRPV5 calcium channels (transient receptor potential vanilloid, type 5), which we discovered earlier in human T cells of the Jurkat line. Immunofluorescence analysis of cells showed the near-membrane localization of TRPV5 proteins and their colocalization with lipid rafts. Membrane cholesterol reduction with the use of methyl β-cyclodextrin (MbCD) led to a decrease in surface exposure of the channels and their diffuse distribution in the cell cytoplasm. An analysis of cell images obtained by immunoelectron microscopy revealed local clusters of TRPV5 proteins in the form of clusters in the plasma membrane of cells. Extraction of membrane cholesterol and destruction of lipid rafts led to the disappearance of channel clusters and the departure of TRPV5 channels from the plasma membrane of cells into the cytoplasm. On the whole, the results showed that the localization of TRPV5 calcium channels in the form of clusters in the plasma membrane is critically dependent on the level of cholesterol and the integrity of lipid rafts in Jurkat T cells.