anti drd2 rabbit polyclonal antibody  (Proteintech)

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

Journal: International Journal of Neuropsychopharmacology

Article Title: Dopamine Receptor D2 and Associated microRNAs Are Involved in Stress Susceptibility and Resistance to Escitalopram Treatment

doi: 10.1093/ijnp/pyv025

Figure Lengend Snippet: Sequences for Primers or miRNAs

Article Snippet: The rabbit polyclonal anti-DRD2 antibody was purchased from Santa Cruz Biotechnology (San Diego, CA).

Techniques: Quantitative RT-PCR

Journal: International Journal of Neuropsychopharmacology

Article Title: Dopamine Receptor D2 and Associated microRNAs Are Involved in Stress Susceptibility and Resistance to Escitalopram Treatment

doi: 10.1093/ijnp/pyv025

Figure Lengend Snippet: Dopamine receptor D2 (DRD2) protein and mRNA expression in the nucleus accumbens (NAc) and striatum. Comparison of DRD2 mRNA (A) and protein (B) expression in the NAc among groups. Comparison of DRD2 mRNA (C) and protein (D) expression in the striatum among groups. * P <.05 vs normal control (NOR) for saline treatment; Δ P <.05 vs chronic unpredictable stress (CUS) for saline treatment; ǂ P <.05 vs maternal deprivation (MD) for saline treatment. E, escitalopamine; S, saline.

Article Snippet: The rabbit polyclonal anti-DRD2 antibody was purchased from Santa Cruz Biotechnology (San Diego, CA).

Techniques: Expressing, Comparison, Control, Saline

Journal: International Journal of Neuropsychopharmacology

Article Title: Dopamine Receptor D2 and Associated microRNAs Are Involved in Stress Susceptibility and Resistance to Escitalopram Treatment

doi: 10.1093/ijnp/pyv025

Figure Lengend Snippet: Correlations between Behavioral Index and Gene Expression

Article Snippet: The rabbit polyclonal anti-DRD2 antibody was purchased from Santa Cruz Biotechnology (San Diego, CA).

Techniques:

Journal: International Journal of Neuropsychopharmacology

Article Title: Dopamine Receptor D2 and Associated microRNAs Are Involved in Stress Susceptibility and Resistance to Escitalopram Treatment

doi: 10.1093/ijnp/pyv025

Figure Lengend Snippet: microRNAs (miRNAs) regulated dopamine receptor D2 (DRD2) expression in vitro. (A) Nucleotide base of DRD2 mRNA 3’ untranslated region (3’-UTR) pairs with miR-9 and miR-326. (B) The expression of FAM (green) after transfection of miR-9 and miR-326 into SH-SY5Y cell. (C) The expression of miR-9 and miR-326 after transfection of mimics or inhibitor into SH-SY5Y cell. (D) The expression of DRD2 mRNA after transfection of mimics or inhibitor into SH-SY5Y cell. (E) Western blot of DRD2 and ß-actin after transfection of mimics and inhibitor into SH-SY5Y cell. (F) Relative luciferase activity of DRD2 (wild-type, WT) 3’UTR and its mutation (MUT) reporter after cotransfection of plasmid and miR-9 mimics (or negative control mimics) into 293T cell. * P < .05.

Article Snippet: The rabbit polyclonal anti-DRD2 antibody was purchased from Santa Cruz Biotechnology (San Diego, CA).

Techniques: Expressing, In Vitro, Transfection, Western Blot, Luciferase, Activity Assay, Mutagenesis, Cotransfection, Plasmid Preparation, Negative Control