The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/500. Detects a band of approximately 48-53 kDa (predicted molecular weight: 48 kDa).
Use at an assay dependent concentration. PubMed: 22155235
FunctionDopamine receptor responsible for neuronal signaling in the mesolimbic system of the brain, an area of the brain that regulates emotion and complex behavior. Its activity is mediated by G proteins which inhibit adenylyl cyclase.
Sequence similaritiesBelongs to the G-protein coupled receptor 1 family.
Post-translational modificationsPolyubiquitinated by the BCR(KLHL12) E3 ubiquitin ligase complex: polyubiquitination does not lead to degradation of DRD4 protein.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dopamine Receptor D4 antibody (ab20424)Image from Wang Q et al., Biochem Biophys Res Commun. 2012;417(1):197-203. Fig 1.; doi: 10.1016/j.bbrc.2011.11.084 with permission from Elsevier.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat gastric tissue labelling Dopamine Receptor D4 with ab20424 at 1/1000. Dopamine Receptor D4 was not found in the gastric mucosa (e - left) but was observed in the gastric enteric plexus (f - right). Paraffin sections were baked at 60°C for 2 hours, then deparaffinized in xylene for 20 minutes. Slides were rehydrated in descending concentrations of alcohol and transferred from 70% alcohol solution to distilled water for rinsing. Before staining, slides were pretreated with 10 mM citric acid (pH 6.0) at 92–98°C for 10 min and incubated in 10% normal serum and 0.1% Triton X-100 for 30 min. The sections were then incubated with the primary antibody, followed by the appropriate secondary antibody. Sections were washed in PBS and mounted in 90% glycerin PBS.
ab20424 stained PC12 cells. The cells were 4% formaldhyde fixed for 10 minutes at room temperature and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1hour at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab20424 at 1/100) overnight at +4°C. The secondary antibody (pseudo-colored green) was Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) used at a 1/1000 dilution for 1hour at room temperature. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.
References for Anti-Dopamine Receptor D4 antibody (ab20424)
This product has been referenced in:
Wang Q et al. Cellular localization of dopamine receptors in the gastric mucosa of rats. Biochem Biophys Res Commun417:197-203 (2012).
Read more (PubMed: 22155235) »
Krishnan B et al. Dopamine receptor mechanisms mediate corticotropin-releasing factor-induced long-term potentiation in the rat amygdala following cocaine withdrawal. Eur J Neurosci31:1027-42 (2010).
Read more (PubMed: 20377617) »