Fig 2: Immunohistology (IH) using an antibody with a DAT C-terminal immunogens. IH was performed using wild-type (WT) mouse tissue, DAT-knock-out (DKO) mouse tissue, or rat tissue in which one hemisphere was lesioned with 6-OHDA to kill dopamine neurons, thus eliminating the possibility of a DAT antibody to produce a signal on the lesioned side (internal negative control). Concentration of DAT antibody indicated in the figure. A, Anti-DAT antibody 431-DATC displaying a faint signal in WT mouse tissue at a concentration of 1:100 and 1:200 and no signal in rat tissue at either concentration. B, Anti-TH antibody in rat tissue used to demonstrate good detection of the 6-OHDA-lesioned side. Scale bar = 6 mm

Fig 3: Western blots depicting the remaining anti-DAT antibodies with N-terminal immunogens. All western blots were performed using mouse striatum samples from DAT-knock-outs (DKO), and wild-type (WT) mice. Total cell lysates were also obtained and utilized from HEK293 and DAT-expressing HEK293 cells. Molecular ladder is indicated beside each figure, with detection of the dopamine transporter (DAT) indicated by a black arrow. Western blots include: (A) anti-DAT antibody PT-22524-1-AP depicting a strong signal with nonspecific binding on a homemade 10% Tris-Glycin acrylamide gel using total cell lysate, (B) anti-DAT antibody PT-22524-1-AP depicting strong nonspecific signal on a homemade 10% Tris-Glycin acrylamide gel using a mouse striatum sample, (C) anti-DAT antibody D6944 depicting a strong DAT signal with some nonspecific binding on a homemade 10% Tris-Glycin acrylamide gel using mouse striatum sample, and (D) anti-DAT antibody MA5-24795 depicting a strong DAT signal with minor nonspecific binding on a Novex 10% Tris-Glycine gel using mouse striatum sample.

Fig 4: Western blots depicting anti-DAT antibodies with 2nd extracellular loop immunogens. All western blots were performed using mouse striatum samples from DAT-knock-outs (DKO), and wild-type (WT) mice. Molecular ladder is indicated beside each figure, with detection of the dopamine transporter (DAT) indicated by a black arrow. Western blots using antibodies 2nd loop immunogens include: (A) anti-DAT antibody SC-32259 depicting no DAT signal and weak nonspecific binding on a Novex 10% Tris-Glycine gel using mouse striatum sample, (B) anti-DAT antibody SC-58517 depicting no DAT signal on a Novex 10% Tris-Glycine gel using mouse striatum sample, and (C) anti-DAT antibody 434-DATEL2 depicting no DAT signal and some strong nonspecific binding on a Novex 10% Tris-Glycine gel using mouse striatum sample.

Fig 5: Western blots depicting anti-DAT antibodies with N-terminal immunogens. All western blots were performed using mouse striatum samples from DAT-knock-outs (DKO), and wild-type (WT) mice. Molecular ladder is indicated beside each figure, with detection of the dopamine transporter (DAT) indicated by a black arrow. Western blots include: (A) anti-DAT antibody MAB369 depicting no signal run on a Novex 4–20% Tris-Glycine gel using mouse striatum sample, (B) anti-DAT antibody AB2231 depicting strong nonspecific signal run on a Novex 4–20% Tris-Glycine gel using mouse striatum sample, (C) anti-DAT antibody SC-32258 depicting a moderate signal with some nonspecific bands run on a Novex 10% Tris-Glycine gel using mouse striatum sample, (D) anti-DAT antibody GTX30992 depicting a weak signal run on a homemade 10% Tris-Glycin acrylamide gel using mouse striatum sample, and (E) anti-DAT antibody ZRB1525 depicting a weak signal run on a Novex 10% Tris-Glycine gel using mouse striatum sample.