rabbit polyclonal d2s primary antibody  (OriGene)

Journal: International Journal of Molecular Sciences

Article Title: Single and Binge Methamphetamine Administrations Have Different Effects on the Levels of Dopamine D2 Autoreceptor and Dopamine Transporter in Rat Striatum

doi: 10.3390/ijms15045884

Figure Lengend Snippet: Dopamine D2 receptor and dopamine transporter (DAT) species in rat striatal synaptosomes. ( A ) The immunoreactivity of the antibody recognizing both D2S and D2L isoforms of D2 receptor; ( B ) the immunoreactivity of the antibody recognizing the D2L receptor; and ( C ) the immunoreactivity of the antibody recognizing the DAT. Abbreviations: D2L, D2long receptor; D2S, D2short receptor; H, homogenate; M, membrane synaptosomal fraction; T, total synaptosomal fraction; V, vesicular synaptosomal fraction.

Article Snippet: Dynabeads (Life Technologies, Grand Island, NY, USA) were incubated in the presence of 2 μL of rabbit polyclonal D2 receptor primary antibody (AB5084P, 1:1000; EDM Millipore), polyclonal DAT primary antibody (SC1433; Santa Cruz Biotechnology Inc.), or radioimmunoprecipitation assay (RIPA) buffer (negative control) for 12 h at 4 °C, followed by the addition of synaptosomal fractions (200 μg of proteins in 100 μL of RIPA buffer) and second incubation (12 h at 4 °C).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Single and Binge Methamphetamine Administrations Have Different Effects on the Levels of Dopamine D2 Autoreceptor and Dopamine Transporter in Rat Striatum

doi: 10.3390/ijms15045884

Figure Lengend Snippet: The effect of binge intraperitoneal (i.p.) METH on the immunoreactivity of pre-synaptic dopamine D2 autoreceptor (D2S) in striatal synaptosomal fractions. Binge METH (4 × 10 mg/kg, every 2 h, i.p.) increased D2 receptor immunoreactivity at 1 h in total synaptosomal fraction (+32%, * p < 0.05) and decreased D2 receptor immunoreactivity at 24 h in all three fractions, total, membrane-endosomal, and cytosol-vesicular fraction (−34% ( ** p < 0.01), −28%, and −11%, respectively). A trend for an increase in D2S immunoreactivity was observed in cytosol-vesicular fraction (+48%, p = 0.069). Abbreviations: M, membrane fraction; METH, methamphetamine; PonS, Ponceau S; SAL, saline; T, total fraction; V, vesicular fraction.

Article Snippet: Dynabeads (Life Technologies, Grand Island, NY, USA) were incubated in the presence of 2 μL of rabbit polyclonal D2 receptor primary antibody (AB5084P, 1:1000; EDM Millipore), polyclonal DAT primary antibody (SC1433; Santa Cruz Biotechnology Inc.), or radioimmunoprecipitation assay (RIPA) buffer (negative control) for 12 h at 4 °C, followed by the addition of synaptosomal fractions (200 μg of proteins in 100 μL of RIPA buffer) and second incubation (12 h at 4 °C).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Single and Binge Methamphetamine Administrations Have Different Effects on the Levels of Dopamine D2 Autoreceptor and Dopamine Transporter in Rat Striatum

doi: 10.3390/ijms15045884

Figure Lengend Snippet: The assessment of dopamine pre-synaptic D2 autoreceptor-dopamine transporter (D2S-DAT) interactions after single and binge METH. Synaptosomes from saline- and METH-treated rats were subjected to co-immunoprecipitation using antibodies against DAT or D2 receptor (D2R). ( A ) Single METH (6 mg/kg, i.v.) did not cause any apparent changes to the levels of D2S-DAT complex (lane 3 vs. 4); ( B ) binge METH (4 × 10 mg/kg, every 2 h, i.p.) increased the levels of D2S-DAT complex (top: lane 2 vs. 4; bottom: lane 3 vs. 4). Controls: IP Control—negative control, Synaptosome—positive control. Abbreviations: IP, immunoprecipitation; i.p., intraperitoneal; i.v., intravenous; METH, methamphetamine; SAL, saline; WB, western blotting.

Article Snippet: Dynabeads (Life Technologies, Grand Island, NY, USA) were incubated in the presence of 2 μL of rabbit polyclonal D2 receptor primary antibody (AB5084P, 1:1000; EDM Millipore), polyclonal DAT primary antibody (SC1433; Santa Cruz Biotechnology Inc.), or radioimmunoprecipitation assay (RIPA) buffer (negative control) for 12 h at 4 °C, followed by the addition of synaptosomal fractions (200 μg of proteins in 100 μL of RIPA buffer) and second incubation (12 h at 4 °C).

Techniques: Immunoprecipitation, Negative Control, Positive Control, Western Blot