Dopamine Transporter (DAT)

The dopamine transporter (DAT) is a 70-80kD member of the Na + -neurotransmitter symporter family of transmembrane (TM) proteins. DAT plays a crucial role in the synaptic clearance of dopamine (DA). It mediates the uptake of dopamine by the presynaptic terminal, thus limiting the strength of the dopamingeric response. One molecule of DA is accompanied by two Na + and one Cl-ion. Molecules such as amphetamine both competitively inhibit DA uptake, and induce DA release through the DAT, increasing the rewarding property of DA. Human DAT is a 620aa, 12 TM phosphoglycoprotein with an N- and C-terminal cytoplasmic domain. It exists as a disulfide-linked oligomer on the cell surface. Phosphorylation of the N-terminus (S7/12) promotes DA release. The C-terminus binds CaMKII, as well as Hic5, Pick1, and synuclein which regulate receptor trafficking and expression. There is an extended 71aa extracellular (EC) loop between TM segments 3 and 4. Glycosylation at this site is necessary for oligomer expression on the cell membrane. The human 42aa DAT C-terminus is 93% aa identical to the mouse DAT C-terminus. The human EC loop is also 93% aa identical to the rhesus monkey EC loop. Applications: Suitable for use in Western Blot and Immunohistochemistry. Other applications not tested. Recommended Dilution: Western Blot: 1:1000. Sufficient for 10 mini-blots. Immunohistochemistry: 1:1000. Frozen sections Optimal dilutions to be determined by the researcher. Storage and Stability: May be stored at 4 degrees C for short-term only. For long-term storage and to avoid repeated freezing and thawing, aliquot and store at -20 degrees C. Aliquots are stable for at least 12 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.