rabbit drd2 polyclonal  (Proteintech)

Journal: PLoS ONE

Article Title: The Effect of Chronic Antipsychotic Drug on Hypothalamic Expression of Neural Nitric Oxide Synthase and Dopamine D2 Receptor in the Male Rat

doi: 10.1371/journal.pone.0033247

Figure Lengend Snippet: Haloperidol and risperidone increased DRD2 mRNA expression in both the MPOA and the PVN. Data were relative to β-actin mRNA. * p <0.05 vs Vehicle.

Article Snippet: The membranes were treated with a blocking buffer (5% nonfat dry milk with 0.1% Tween-20) and incubated with primary nNOS and DRD2 antibody respectively (mouse monoclonal antibody against nNOS 1∶500, Transduction Laboratories, Lexington, KY; Rabbit polyclonal antibody against DRD2 1:200, Chemicon International Inc., Temecula, CA) at 4°C overnight.

Techniques: Expressing

Journal: PLoS ONE

Article Title: The Effect of Chronic Antipsychotic Drug on Hypothalamic Expression of Neural Nitric Oxide Synthase and Dopamine D2 Receptor in the Male Rat

doi: 10.1371/journal.pone.0033247

Figure Lengend Snippet: The calculated ODs of nNOS (B) and DRD2 (D) protein bands were normalized to each β-tubulin OD and expressed as a ratio. nNOS protien in the MPOA but not the PVN was significantly decreased by haloperidol. Haloperidol and risperidone increased DRD2 protien expression in both the MPOA and the PVN. Lanes: 1 = Vehicle; 2 = haloperidol; 3 = risperdone; 4 = quetiapine. Data are means ± s.e.means and expressed in arbitrary units. * p <0.05 vs Vehicle; ** p <0.001 vs Vehicle.

Article Snippet: The membranes were treated with a blocking buffer (5% nonfat dry milk with 0.1% Tween-20) and incubated with primary nNOS and DRD2 antibody respectively (mouse monoclonal antibody against nNOS 1∶500, Transduction Laboratories, Lexington, KY; Rabbit polyclonal antibody against DRD2 1:200, Chemicon International Inc., Temecula, CA) at 4°C overnight.

Techniques: Expressing

Journal: ACS Chemical Neuroscience

Article Title: Developing Photoaffinity Probes for Dopamine Receptor D 2 to Determine Targets of Parkinson’s Disease Drugs

doi: 10.1021/acschemneuro.2c00544

Figure Lengend Snippet: Intracellular calcium flux assay. HEK293T cell line stably expressing constructs for human DRD2 and a chimeric G-protein is loaded with calcium-sensing dye, Fura-4. After dosing the probe, confocal microscopy is used to determine the calcium flux in the cell by change in dye fluorescence. EC 50 curves determined with GraphPad software using a Hill slope of 1.0.

Article Snippet: Following blocking, the membrane was incubated with either 1 μg/mL Anti-Strep Tag II rabbit polyclonal antibody (Abcam, ab76949) or 2 μg/mL Anti-DRD2 rabbit polyclonal antibody (AbClonal, A12930) in TBST with 5% BSA overnight at 4 °C.

Techniques: Calcium Flux Assay, Stable Transfection, Expressing, Construct, Confocal Microscopy, Fluorescence, Software

Journal: ACS Chemical Neuroscience

Article Title: Developing Photoaffinity Probes for Dopamine Receptor D 2 to Determine Targets of Parkinson’s Disease Drugs

doi: 10.1021/acschemneuro.2c00544

Figure Lengend Snippet: β-Arrestin recruitment analysis. (a) PRESTO-TANGO assay schematic: a ligand binds a chimeric DRD2 receptor, which then recruits β-arrestin fused with a TEV protease. The protease cuts a site between the receptor and a fused transcription factor, which then transits to the nuclease to initiate transcription of a luciferase gene. The luciferase activity is subsequently quantified. (b) Agonism of β-arrestin recruitment is quantified in EC 50 curves via the detection of luciferase activity, using a Hill slope of 1.0.

Article Snippet: Following blocking, the membrane was incubated with either 1 μg/mL Anti-Strep Tag II rabbit polyclonal antibody (Abcam, ab76949) or 2 μg/mL Anti-DRD2 rabbit polyclonal antibody (AbClonal, A12930) in TBST with 5% BSA overnight at 4 °C.

Techniques: Luciferase, Activity Assay

Journal: ACS Chemical Neuroscience

Article Title: Developing Photoaffinity Probes for Dopamine Receptor D 2 to Determine Targets of Parkinson’s Disease Drugs

doi: 10.1021/acschemneuro.2c00544

Figure Lengend Snippet: Photo-cross-linking of dye-clicked probe: confocal microscopy. (a) Schematic of the methodology used in the labeling process. Cells stably expressing DRD2 fused to an N-terminal Strep Tag II are treated with DRD2-targeting probes 5 or 7 at 5 μM, photo-cross-linked, and excess probe is washed out. An Alexa Fluor 555 azide is then clicked to the probe, washed out, and cells are treated with an anti-Strep-Tag II antibody and fluorescent secondary to visualize DRD2. Nuclei were stained with DAPI. (b) Confocal microscopy results of labeled cells. All probes show some degree of labeling. However, probes 5 and 7 show a notable increase in the labeling density. Images taken with 40× magnification, scale bar: 10 μm.

Article Snippet: Following blocking, the membrane was incubated with either 1 μg/mL Anti-Strep Tag II rabbit polyclonal antibody (Abcam, ab76949) or 2 μg/mL Anti-DRD2 rabbit polyclonal antibody (AbClonal, A12930) in TBST with 5% BSA overnight at 4 °C.

Techniques: Confocal Microscopy, Labeling, Stable Transfection, Expressing, Strep-tag, Staining

Journal: ACS Chemical Neuroscience

Article Title: Developing Photoaffinity Probes for Dopamine Receptor D 2 to Determine Targets of Parkinson’s Disease Drugs

doi: 10.1021/acschemneuro.2c00544

Figure Lengend Snippet: Flow cytometry quantification of probe labeling. (a) Schematic of flow cytometry workflow. (b) DRD2-expressing 293T cells or untransduced 293T cells (negative control) are treated with a 100 nm probe, which is photo-cross-linked, and an Alexa Fluor 555 azide is then “clicked” onto the probe. The cells were then analyzed by flow cytometry. P values determined using two-way ANOVA in GraphPad; **** corresponds to P < 0.0001.

Article Snippet: Following blocking, the membrane was incubated with either 1 μg/mL Anti-Strep Tag II rabbit polyclonal antibody (Abcam, ab76949) or 2 μg/mL Anti-DRD2 rabbit polyclonal antibody (AbClonal, A12930) in TBST with 5% BSA overnight at 4 °C.

Techniques: Flow Cytometry, Labeling, Expressing, Negative Control

Journal: ACS Chemical Neuroscience

Article Title: Developing Photoaffinity Probes for Dopamine Receptor D 2 to Determine Targets of Parkinson’s Disease Drugs

doi: 10.1021/acschemneuro.2c00544

Figure Lengend Snippet: Photo-cross-linking of probes 5 and 7 to DRD2 visualized with Western blot. Lanes 1–5 correspond to samples with probe 5 at 100 nm, lanes 6–10 correspond to samples treated with probe 7 at 100 nm, and lanes 11–14 correspond to samples treated with negative control probe 16 . Lanes: 1, 6, and 11 are the anti-DRD2 antibody channel, lanes 2, 3, 7, 8, and 12 are fluorescence of the Alexa Fluor 555 clicked to the probes 5 , 7 , or 16 , lanes 4, 9, and 13 are the anti-Strep Tag antibody channel, and lanes 5, 10, and 14 are the overlaid channels for the respective probes.

Article Snippet: Following blocking, the membrane was incubated with either 1 μg/mL Anti-Strep Tag II rabbit polyclonal antibody (Abcam, ab76949) or 2 μg/mL Anti-DRD2 rabbit polyclonal antibody (AbClonal, A12930) in TBST with 5% BSA overnight at 4 °C.

Techniques: Western Blot, Negative Control, Fluorescence, Strep-tag

Journal: Molecular Neurobiology

Article Title: Electroacupuncture Alleviates Anxiety-Like Behaviors Induced by Chronic Neuropathic Pain via Regulating Different Dopamine Receptors of the Basolateral Amygdala

doi: 10.1007/s12035-022-02911-6

Figure Lengend Snippet: Expression of different DRs and EA-mediated regulation of these receptors in SNI mice. A and B Western blot images showing the protein levels and the quantification of DRD1 ( A ) and DRD2 ( B ) in the AMY 14 days after SNI surgery. ( n = 5–6 per group). C and D Western blot images showing the protein levels and the quantification of DRD1 ( C ) and DRD2 ( D ) in the AMY of SNI mice treated with EA. ( n = 6 per group). Data are expressed as the mean ± SEM. * p < 0.05 comparison of the two groups

Article Snippet: The membranes were blocked with 5% nonfat milk dissolved in TBST at room temperature for 1 h and incubated with the primary antibodies anti-DRD1 rabbit polyclonal antibody (1:1000, ab81296, Abcam) and anti-DRD2 rabbit polyclonal antibody (1:500, 55,084–1-AP, Proteintech) overnight at 4 °C.

Techniques: Expressing, Western Blot

Journal: Molecular Neurobiology

Article Title: Electroacupuncture Alleviates Anxiety-Like Behaviors Induced by Chronic Neuropathic Pain via Regulating Different Dopamine Receptors of the Basolateral Amygdala

doi: 10.1007/s12035-022-02911-6

Figure Lengend Snippet: The effects of EA and the DRD2 antagonist sulpiride on anxiety-like behaviors induced by microinjection of the DRD2 agonist quinpirole in the BLA. A Schematic of EA treatment and drug injection and behavioral testing. B Diagram showing the cannulation location (left) and a representative coronal section of the BLA from a mouse brain showing the cannula tip upon injection (right). C The effect of treatment in each group on the PWTs. ( n = 9 per group). D – F The effect of treatment in each group on anxiety-like behaviors in the EPM ( D ) and OFT ( E – F ). ( n = 7–9 per group). G Representative diagram showing tracked movement and activity heatmaps from the EPM. H Representative diagram showing tracked movement and activity heatmaps from the OFT. Data are expressed as the mean ± SEM. Tukey’s post hoc test: * p < 0.05 comparison of the two groups

Article Snippet: The membranes were blocked with 5% nonfat milk dissolved in TBST at room temperature for 1 h and incubated with the primary antibodies anti-DRD1 rabbit polyclonal antibody (1:1000, ab81296, Abcam) and anti-DRD2 rabbit polyclonal antibody (1:500, 55,084–1-AP, Proteintech) overnight at 4 °C.

Techniques: Injection, Activity Assay

Journal: Molecular Neurobiology

Article Title: Electroacupuncture Alleviates Anxiety-Like Behaviors Induced by Chronic Neuropathic Pain via Regulating Different Dopamine Receptors of the Basolateral Amygdala

doi: 10.1007/s12035-022-02911-6

Figure Lengend Snippet: The effects of EA and the DRD2 antagonist sulpiride on pain-related allodynia and anxiety-like behaviors induced by SNI in mice. A Schematic of EA treatment and drug injection and behavioral testing. B Diagram showing the cannulation location (left) and a representative coronal section of the BLA from a mouse brain containing a cannula tip after injection (right). C The effect of treatment in each group on the PWTs. ( n = 9–10 per group). D – F The effect of treatment in each group on anxiety-like behaviors in the EPM ( D ) and OFT ( E – F ). ( n = 7–10 per group). G Representative diagram showing tracked movement and activity heatmaps from the EPM. H Representative diagram showing tracked movement and activity heatmaps from the OFT. Data are expressed as the mean ± SEM. Tukey’s post hoc test: + p < 0.05 compared with the sham + vehicle group; # p < 0.05 compared with the SNI + vehicle group; $ p < 0.05 compared with the SNI + sulpiride group; * p < 0.05 comparison between the two groups

Article Snippet: The membranes were blocked with 5% nonfat milk dissolved in TBST at room temperature for 1 h and incubated with the primary antibodies anti-DRD1 rabbit polyclonal antibody (1:1000, ab81296, Abcam) and anti-DRD2 rabbit polyclonal antibody (1:500, 55,084–1-AP, Proteintech) overnight at 4 °C.

Techniques: Injection, Activity 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: 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