rabbit polyclonal d2s primary antibody  (OriGene)

Journal: Biomedicines

Article Title: Dopamine D2 Long Receptors Are Critical for Caveolae-Mediated α-Synuclein Uptake in Cultured Dopaminergic Neurons

doi: 10.3390/biomedicines9010049

Figure Lengend Snippet: The primary sequence of mouse dopamine D2 receptor long ( left ) and short ( right ) isoforms. Amino acid sequences in red letters are specifically expressed in the long type. The dopamine D2 receptor short isoform ( right ) lacks these 29 amino acid sequences, which interact with fatty acid-binding protein 3 (FABP3). Asterisks indicate the antigen used to produce D 2L specific antibody used in this study.

Article Snippet: Following fixation with 4% paraformaldehyde for 30 min, the cells were incubated with 0.1% Triton X-100 for 15 min. After pre-blocking with 5% goat serum in phosphate-buffered saline (PBS) for 1 h, they were incubated overnight at 4 °C with the following primary antibodies: rabbit anti-TH affinity-purified polyclonal antibody (1:400; Millipore, AB152, Billerica, MA, USA), mouse anti-TH monoclonal antibody (1:200; Millipore, MAB318), mouse anti-human FABP3 monoclonal antibody, clone 66E2 (1:50; Hycult Biotech, HM2016, Uden, Netherlands), rabbit anti-dopamine D2 receptor (DRD2) polyclonal antibody (1:500; Proteintech, 55084-l-AP, Rosemont, IL, USA), and rabbit anti-caveolin-1 polyclonal antibody (1:500; Abcam, ab2910, Cambridge, UK).

Techniques: Sequencing, Binding Assay

Journal: Biomedicines

Article Title: Dopamine D2 Long Receptors Are Critical for Caveolae-Mediated α-Synuclein Uptake in Cultured Dopaminergic Neurons

doi: 10.3390/biomedicines9010049

Figure Lengend Snippet: Cultured dopaminergic neurons require dopamine D2 receptors to take up α-synuclein. ( A ) Representative images of TH + mesencephalic neurons at 12 days in vitro (DIV) derived from wild type (WT) or D 2L −/− C57BL6 mice. Neurons were treated with 1 μM ATTO-550-labeled α-synuclein monomer for 48 h and stained with anti-TH antibody (TH, green). The magnified images were enlarged by three times. Scale bar 10 μm. The right graph shows the quantitative analysis of ATTO-550-labeled α-synuclein monomer fluorescence intensity of individual TH + neurons. **** p < 0.0001 in wild type (WT) versus D 2L −/− , n = 34 in three independent experiments. ( B ) Representative images of TH + mesencephalic neurons derived from wild type or D2 null knockout mice. Neurons were treated with ATTO-550-labeled α-synuclein monomer in the same condition as in ( A ) and stained with anti-TH antibody (TH, green) and dopamine D2 receptor (DRD2, blue). The magnified images were enlarged by three times. Scale bar 10 μm. The quantitative analysis of ATTO-550-labeled α-synuclein monomer fluorescence intensity of individual TH + neurons on the right. **** p < 0.0001 in WT versus D2 null knockout (D2 null), n = 28 in three independent experiments.

Article Snippet: Following fixation with 4% paraformaldehyde for 30 min, the cells were incubated with 0.1% Triton X-100 for 15 min. After pre-blocking with 5% goat serum in phosphate-buffered saline (PBS) for 1 h, they were incubated overnight at 4 °C with the following primary antibodies: rabbit anti-TH affinity-purified polyclonal antibody (1:400; Millipore, AB152, Billerica, MA, USA), mouse anti-TH monoclonal antibody (1:200; Millipore, MAB318), mouse anti-human FABP3 monoclonal antibody, clone 66E2 (1:50; Hycult Biotech, HM2016, Uden, Netherlands), rabbit anti-dopamine D2 receptor (DRD2) polyclonal antibody (1:500; Proteintech, 55084-l-AP, Rosemont, IL, USA), and rabbit anti-caveolin-1 polyclonal antibody (1:500; Abcam, ab2910, Cambridge, UK).

Techniques: Cell Culture, In Vitro, Derivative Assay, Labeling, Staining, Fluorescence, Knock-Out