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Rat anti dat

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The Rat anti-DAT is a laboratory reagent used to detect the dopamine transporter (DAT) protein in rat samples. It is a specific antibody that binds to the DAT protein, allowing its identification and quantification in various experimental applications.

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Lab products found in correlation

Mouse anti tuj1, by Fortrea (3 mentions) Goat anti chat, by Merck Group (2 mentions) Mouse anti th, by Merck Group (2 mentions) Rabbit anti th, by Merck Group (2 mentions) Mouse anti map2, by Merck Group (2 mentions) Rabbit anti tuj1, by Fortrea (2 mentions) Fluorescent tagged secondary antibodies, by Thermo Fisher Scientific (1 mentions) Vectorshield mounting medium, by Vector Laboratories (1 mentions) Rabbit anti eaat3, by Alpha Diagnostic (1 mentions) Mouse anti nr1, by Merck Group (1 mentions) Mouse anti gfap, by Cell Signaling Technology (1 mentions) Mouse anti β actin, by Merck Group (1 mentions) Alexa fluor 488 or 555, by Thermo Fisher Scientific (1 mentions) Rabbit anti nurr1, by Santa Cruz Biotechnology (1 mentions) Rabbit anti pitx3, by Thermo Fisher Scientific (1 mentions) Mouse anti syn1, by Synaptic Systems (1 mentions) Goat anti foxa2, by Santa Cruz Biotechnology (1 mentions) Rabbit anti gapdh, by Cell Signaling Technology (1 mentions) Alexa 488, by Thermo Fisher Scientific (1 mentions) Mouse anti neun, by Merck Group (1 mentions)

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Anti-DAT (7 citations)

7 protocols using rat anti dat

1

Detailed Immunofluorescence Staining Protocol

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Immunofluorescence staining on 35-mm glass-bottomed dishs was described previously [11 (link)]. The primary antibodies were used at dilutions recommended by the manufacturers. The antibodies (anti human in all cases) used were mouse anti-Tuj1 (Covance, 1:800), rabbit anti-TH (Pel-Freez, 1:1 000), rat anti-DAT (Chemicon, 1:5 000), rabbit anti-DDC (Chemicon, 1:500), rat anti-Serotonin (Chemicon, 1:200), goat anti-ChAT (Chemicon, 1:100), mouse anti-HN (Millipore, 1:200). The immunostaining was developed with appropriate fluorescent-tagged secondary antibodies (Invitrogen, Carlsbad, CA, USA). In all cases, cellular nuclei were stained with DAPI in VectorShield mounting medium (Vector Laboratories, Burlingame, CA, USA). Images were collected with a Ziess or Leica SP5 confocal laser-scanning microscope.
XinJian L., Qian H., Fang L, & Chuan-Yuan L. (2014). Enhancing the efficiency of direct reprogramming of human primary fibroblasts into dopaminergic neuron-like cells through p53 suppression. Science China. Life sciences, 57(9), 867-875.
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2

Multimodal Neurotransmitter Receptor Detection

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Rabbit anti-EAAT3 (Alpha Diagnostics), mouse anti-TH (Sigma), rat anti-DAT (Chemicon), Rabbit anti-EAAT3 (Alpha diagnostics), rabbit anti-EAAT2 (made inhouse), rabbit anti-GluR4 (Millipore) rabbit anti-GluR1 (Upstate), and mouse anti-NR1 (Millipore),
Underhill S.M., Wheeler D.S., Li M., Watts S.D., Ingram S.L, & Amara S.G. (2014). Amphetamine Modulates Glutamatergic Neurotransmission through Endocytosis of the Excitatory Amino Acid Transporter EAAT3 in Dopamine Neurons. Neuron, 83(2), 404-416.
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3

Quantitative Immunohistochemistry of Dopaminergic Markers

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Paraffin-embedded brain blocks were sliced into 5 μm coronal sections and processed for IHC (SN blocks) or hematoxylin-eosin staining (HIPP blocks). Following antigen retrieval and inactivation of endogenous peroxidase activity, SN sections were incubated with mouse anti-TH (1:5000, Sigma) or rat anti-DAT (1:1500, Chemicon) at 4°C overnight. After washing, the sections were respectively incubated with biotinylated goat anti-mouse IgG (1:300) or biotinylated goat anti-rat IgG (1:300) antibodies for 2 h at room temperature. Following 1-h incubation at room temperature in horseradish peroxidase-conjugated streptavidin (1:300), the sections were stained for 5 min in a solution containing 0.05% diaminobenzidine and 0.03% H2O2 in 0.05 M Tris-HCl buffer (pH 7.6). A computer-assisted image analysis system was used to measure AOD for TH and DAT immunoreactivity in the SN and to count the number of HE-stained cells with pyknotic appearance in the hippocampal CA1 region.
Yan W., Zhang T., Kang Y., Zhang G., Ji X., Feng X, & Shi G. (2021). Testosterone ameliorates age-related brain mitochondrial dysfunction. Aging (Albany NY), 13(12), 16229-16247.
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4

Antibody Sources for VMAT2, DAT, and TH

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Rabbit polyclonal anti-VMAT2 serum was raised against a peptide in the C-terminal region of mouse VMAT2 (CTQNNVQPYPVGDDEESESD).35 (link) Rat anti-DAT and rabbit anti-TH were purchased from Millipore. Mouse anti-GFAP was purchased from Cell Signaling Technologies. Mouse anti-β-actin was purchased from Sigma. All secondary antibodies were purchased from Jackson ImmunoResearch Laboratories.
Lohr K.M., Stout K.A., Dunn A.R., Wang M., Salahpour A., Guillot T.S, & Miller G.W. (2015). Increased Vesicular Monoamine Transporter 2 (VMAT2; Slc18a2) Protects against Methamphetamine Toxicity. ACS chemical neuroscience, 6(5), 790-799.
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5

Immunofluorescence Staining of Neuronal Markers

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Cells were fixed with 4% paraformaldehyde for 5 min and permeabilized with 0.2% Triton X-100/PBS. After blocking (4% BSA/1% Cosmic Calf Serum in PBS) for at least 30 min, primary antibodies were added and incubated overnight at 4°C. After three washes, secondary antibody conjugated with Alexa Fluor 488 or 555 (Molecular Probes) was added and incubated for 60 min. The wells were then incubated with 4′,6-diamidino-2-phenylindole (DAPI) nuclear counterstain for 5 min before analysis. Antibodies used were sheep anti-TH (Pel-Freez), mouse anti-EN1 (Development Studies Hybridoma Bank, IA), rabbit anti-PRPH (Millipore), mouse anti-MAP2 (Sigma), mouse anti-TUJ1 (Covance), rabbit anti-TUJ1 (Covance), rabbit anti-PITX3 (Invitrogen), rabbit anti-NURR1 (Santa Cruz Biotechnologies), goat anti-FOXA2 (Santa Cruz), mouse anti-SYN1 (Synaptic Systems), and rat anti-DAT (Millipore). See also Table S2.
Ng Y.H., Chanda S., Janas J.A., Yang N., Kokubu Y., Südhof T.C, & Wernig M. (2021). Efficient generation of dopaminergic induced neuronal cells with midbrain characteristics. Stem Cell Reports, 16(7), 1763-1776.
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6

Neurosphere Differentiation Analysis

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At various time points including day 50 of differentiation for HPLC analysis, the neurospheres were dissociated with ACCUTASE™ into single cells. 1 × 106 cells per vial would be snap frozen in liquid N2 after washing with PBS. Western blot was carried out using 30 μg of total proteins per sample with primary antibodies (1:1000 rat anti-DAT (Millipore, USA), 1:1000 rabbit anti-TH (Pel-Freez, USA) and 1:10,000 rabbit anti-GAPDH (Cell Signaling, USA)). HPLC analysis was performed as described previously [15 (link)].
Goggi J.L., Qiu L., Liao M.C., Khanapur S., Jiang L., Boominathan R., Hartimath S.V., Cheng P., Yong F.F., Soh V., Deng X., Lin Y.M., Haslop A., Tan P.W., Zeng X., Lee J.W., Zhang Z., Sadasivam P., Tan E.K., Luthra S.K., Shingleton W.D., Oh S.K., Zeng L, & Robins E.G. (2020). Dopamine transporter neuroimaging accurately assesses the maturation of dopamine neurons in a preclinical model of Parkinson’s disease. Stem Cell Research & Therapy, 11, 347.
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7

Immunostaining Protocol for Cultured Cells

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Immunostaining for cultured cells was carried out as described previously (Yamamizu et al. 2013 (link)). Briefly, 4% paraformaldehydefixed cells were blocked by 1% skim milk (BioLab) and incubated overnight with primary antibodies at 4°C. For immunofluorescent staining, anti-mouse, rat, rabbit, or goat IgG antibodies conjugated with Alexa488 or Alexa546 (Invitrogen) were used as secondary antibodies. Primary antibodies were as follows: mouse anti-TUJ1 (1:500; Covance, Princeton, NJ); rabbit anti-TUJ1 (1:500; Covance); mouse anti-MAP2 (1:500; Sigma-Aldrich, St. Louis, MO); mouse anti-NEUN (1:100; Millipore); rabbit anti-TH (1:500; Millipore); rat anti-DAT (1:500; Millipore); rabbit anti-VMAT2 (1:500; Millipore); rabbit anti-AADC (1:500; Abcam); rabbit anti-ALDH1A1 (1:500; Abcam); rabbit anti-GABA (1:500; Sigma-Aldrich); mouse anti-GAD67 (1:500; Millipore); mouse anti-PV (1:500; Millipore); rat anti-SST (1:500; Millipore); mouse anti-ISL1/ISL2 (1:500; DSHB); mouse anti-Hb9 (1:500; DSHB); goat anti-ChAT (1:500; Sigma-Aldrich). Differentiated neurons were photographed with inverted fluorescent microscopy (Eclipse TE300; Nikon) with the use of NIS-Elements Software (Nikon).
Teratani-Ota Y., Yamamizu K., Piao Y., Sharova L., Amano M., Yu H., Schlessinger D., Ko M.S, & Sharov A.A. (2016). Induction of specific neuron types by overexpression of single transcription factors. In vitro cellular & developmental biology. Animal, 52(9), 961-973.
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