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71241
Dopamine D(2) Receptor/D2R (E1U8K) Rabbit mAb
Primary Antibodies
Monoclonal Antibody

Dopamine D(2) Receptor/D2R (E1U8K) Rabbit mAb #71241

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  1. IF
Immunofluorescence Image 1 - Dopamine D(2) Receptor/D2R (E1U8K) Rabbit mAb

Confocal immunofluorescent analysis of mouse striatum (left, positive) and mouse cerebellum (right, negative) using Dopamine D(2) Receptor/D2R (E1U8K) Rabbit mAb (green). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).

To Purchase # 71241S
Product # Size Price
71241S
100 µl $ 268

Supporting Data

REACTIVITY M
SENSITIVITY Endogenous
MW (kDa)
Source/Isotype Rabbit IgG

Application Key:

  • W-Western
  • IP-Immunoprecipitation
  • IHC-Immunohistochemistry
  • ChIP-Chromatin Immunoprecipitation
  • IF-Immunofluorescence
  • F-Flow Cytometry
  • E-P-ELISA-Peptide

Species Cross-Reactivity Key:

  • H-Human
  • M-Mouse
  • R-Rat
  • Hm-Hamster
  • Mk-Monkey
  • Mi-Mink
  • C-Chicken
  • Dm-D. melanogaster
  • X-Xenopus
  • Z-Zebrafish
  • B-Bovine
  • Dg-Dog
  • Pg-Pig
  • Sc-S. cerevisiae
  • Ce-C. elegans
  • Hr-Horse
  • All-All Species Expected

Product Usage Information

Application Dilution
Immunofluorescence (Frozen) 1:50 - 1:200

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Protocol

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Immunofluorescence (Frozen)

A. Solutions and Reagents

NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.

  1. 20X Phosphate Buffered Saline (PBS): (9808) To prepare 1L 1X PBS: add 50 ml 20X PBS to 950 ml dH2O, mix. Adjust pH to 8.0.
  2. Formaldehyde: 16%, methanol free, Polysciences, Inc. (cat# 18814), use fresh and store opened vials at 4°C in dark, dilute in 1X PBS for use.
  3. Blocking Buffer: (1X PBS / 5% normal serum / 0.3% Triton™ X-100): To prepare 10 ml, add 0.5 ml normal serum from the same species as the secondary antibody (e.g., Normal Goat Serum (#5425) to 9.5 ml 1X PBS) and mix well. While stirring, add 30 µl Triton™ X-100.
  4. Antibody Dilution Buffer: (1X PBS / 1% BSA / 0.3% Triton™ X-100): To prepare 10 ml, add 30 µl Triton™ X-100 to 10 ml 1X PBS. Mix well then add 0.1 g BSA (#9998), mix.
  5. Recommended Fluorochrome-conjugated Anti-Rabbit secondary antibodies:

    NOTE: When using any primary or fluorochrome-conjugated secondary antibody for the first time, titrate the antibody to determine which dilution allows for the strongest specific signal with the least background for your sample.

  6. Prolong® Gold AntiFade Reagent (#9071), Prolong® Gold AntiFade Reagent with DAPI (#8961).

B. Specimen Preparation - Frozen/Cryostat Sections (IF-F)

  1. For fixed frozen tissue proceed with Immunostaining (Section C).
  2. For fresh, unfixed frozen tissue, please fix immediately, as follows:
    1. Cover sections with 4% formaldehyde dilute in 1X PBS.

      NOTE: Formaldehyde is toxic, use only in fume hood.

    2. Allow sections to fix for 15 minutes at room temperature.
    3. Aspirate liquid, rinse three times in 1X PBS for 5 minutes each.
    4. Proceed with Immunostaining (Section C).

C. Immunostaining

NOTE: All subsequent incubations should be carried out at room temperature unless otherwise noted in a humid light-tight box or covered dish/plate to prevent drying and fluorochrome fading.

  1. Block specimen in Blocking Buffer for 60 min.
  2. While blocking, prepare primary antibody by diluting as indicated on protocol on product webpage in Antibody Dilution Buffer.
  3. Aspirate blocking solution, apply diluted primary antibody.
  4. Incubate overnight at 4°C.
  5. Rinse three times in 1X PBS for 5 min each.
  6. Incubate specimen in fluorochrome-conjugated secondary antibody diluted in Antibody Dilution Buffer for 1–2 hr at room temperature in the dark.
  7. Rinse three times in 1X PBS for 5 min each.
  8. Coverslip slides with Prolong® Gold Antifade Reagent (#9071) or Prolong® Gold Antifade Reagent with DAPI (#8961).
  9. For best results, allow mountant to cure overnight at room temperature. For long-term storage, store slides flat at 4°C protected from light.

posted November 2006

revised July 2016

Protocol Id: 151

Specificity / Sensitivity

Dopamine D(2) Receptor/D2R (E1U8K) Rabbit mAb recognizes endogenous levels of total dopamine D(2) receptor/D2R protein. Signal in the lamina propria of mouse small intestine was observed by Immunofluorescence and is presumed to be non-specific.

Species Reactivity:

Mouse

Species predicted to react based on 100% sequence homology:

Human, Rat

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro300 of human dopamine D(2) receptor/D2R protein.

Background

Dopamine is a neurotransmitter that plays important roles in the brain, particularly in dopamine pathways that control the motivational component of reward-motivated behavior. These behavioral outputs are generated by the basal ganglia via its interaction with multiple brain areas that modulate sensorimotor, emotional, and cognitive information (1). The brain’s major dopaminergic input is into the striatum, a region of the basal ganglia composed of GABAergic medium spiny neurons (MSN). Two major subpopulations of MSN exist in the striatum that are distinguished by the expression of dopamine receptor subtypes, the dopamine D(1) receptor subtype and the dopamine D(2) receptor subtype (D1R and D2R, respectively) (2,3). As a family of proteins, dopamine receptors are a class of G-protein coupled-receptors (GPCRs) consisting of 5 subtypes that, upon initiation, drive downstream signaling cascades that modulate neuronal function (1). Dopamine receptors form homo- and hetero-multimers with subunits within their protein family but also with other GPCRs, including Adenosine Receptor A2a, suggesting that dopamine receptor activity might be finely tuned and altered under certain conditions (4). Dopamine receptors have been studied as a therapeutic target for several neuropsychiatric and developmental disorders, as well as neurodegenerative diseases, including Parkinson’s disease (5-8). Dopamine receptors are also expressed outside of the brain and may have diverse functions beyond the central nervous system, including regulating innate and adaptive immunity (9).

  1. Beaulieu, J.M. and Gainetdinov, R.R. (2011) Pharmacol Rev 63, 182-217.
  2. Kebabian, J.W. and Calne, D.B. (1979) Nature 277, 93-6.
  3. Bertran-Gonzalez, J. et al. (2008) J Neurosci 28, 5671-85.
  4. Agnati, L.F. et al. (2016) Rev Neurosci 27, 1-25.
  5. Komatsu, H. et al. (2019) Int J Mol Sci 20, pii: E3207. doi: 10.3390/ijms20133207.
  6. Klein, M.O. et al. (2019) Cell Mol Neurobiol 39, 31-59.
  7. Hurley, M.J. and Jenner, P. (2006) Pharmacol Ther 111, 715-28.
  8. Stocchi, F. et al. (2016) Expert Opin Pharmacother 17, 1889-902.
  9. Vidal, P.M. and Pacheco, R. (2019) J Neuroimmune Pharmacol 19, doi: 10.1007/s11481-019-09834-5.

Pathways & Proteins

Explore pathways + proteins related to this product.

For Research Use Only. Not For Use In Diagnostic Procedures.
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