D1 receptor | knockout validation | Sigma-Aldrich D2994

This is a knockout-validated antibody summary, based on the publication "Evidence against dopamine D1/D2 receptor heteromers", as cited below [1]. The antibody was also validated in the article "Validation of dopamine receptor DRD1 and DRD2 antibodies using receptor deficient mice" for immunohistochemistry and western blot (see figure 1a, 2a) [2]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Antibody information

Monoclonal IgG2a

Company: Sigma-Aldrich (under exclusive license from Emory University)

Antibody: D1 dopamine receptor

Catalog number: D2994 (clone 1-1-F11 s.6)

Summary: raised against a recombinant fusion protein containing the C-terminal 97 amino acids from the human D1 dopamine receptor. It detects monkey, human and rat D1 dopamine receptor. Supplier recommended for immunohistochemistry and Western blotting.

Validation Method

Immunohistochemistry

Sample

Mouse brain (nucleus accumbens) fixed with 4% paraformaldehyde in 0.1M sodium phosphate, pH 7.5

Blocking agent

Tris-buffered saline, pH 7.4 containing 10 fetal bovine serum and 0.5% Triton X-100 for 2 hours at room temperature

Primary incubation

Incubated overnight at 4oC at a dilution of 1:200 - 1:500

Secondary incubation

Alexa 647-conjugated goat anti-rat (Invitrogen) at 1:1000 for 2 hours at room temperature

Detection

Olympus Fluoview FV1000 confocal laser scanning microscope

Figure

See figure 5 from the article [1].

References
  1. Frederick A, Yano H, Trifilieff P, Vishwasrao H, Biezonski D, Meszaros J, et al. Evidence against dopamine D1/D2 receptor heteromers. Mol Psychiatry. 2015;20:1373-85 pubmed publisher
  2. Stojanovic T, Orlova M, Sialana F, Hoger H, Stuchlík S, Milenkovic I, et al. Validation of dopamine receptor DRD1 and DRD2 antibodies using receptor deficient mice.. Amino Acids. 2017;49:1101-1109 pubmed publisher