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Expression in Sf9 insect cells, purification and functional reconstitution of the human proton-coupled folate transporter (PCFT, SLC46A1).

PLoS One.. 2017-05; 
Date SS,Fiori MC,Altenberg GA,Jansen M.
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Catalog Antibody ... (Mountain View, CA). The antibody against the His tag (THE TM Anti-His mAb) was purchased from GenScript (Piscataway, NJ). The tritiated folic acid derivative (folic acid, diammonium salt, [3',5',7,9- 3 H], 19.4 Ci/mmol) was purchased from Moravek Biochemicals Inc. ... Get A Quote

摘要

The proton-coupled folate transporter (PCFT) provides an essential uptake route for the vitamin folic acid (B9) in mammals. In addition, it is currently of high interest for targeting chemotherapeutic agents to tumors due to the increased folic acid requirement of rapidly dividing tumor cells as well as the upregulated PCFT expression in several tumors. To understand its function, determination of its atomic structure and molecular mechanism of transport are essential goals that require large amounts of functional PCFT. Here, we present a high-level heterologous expression system for human PCFT using a recombinant baculovirus and Spodoptera frugiperda (Sf9) insect cells. We demonstrate folate transport function... More

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