Purification notesPurified from rabbit antiserum by affinity chromatography using epitope specific phosphopeptide. The antibody against non-phosphopeptide was removed by chromatography using non-phosphopeptide corresponding to the phosphorylation site.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/50 - 1/100.
Use a concentration of 10 µg/ml.
RelevanceThe opioid receptors are G-protein coupled, seven-transmembrane domain receptors for enkephalins, endorphins, and dynorphins. Three different opioid receptor subtypes (kappa , delta, and mu) were first identified by their different selectivities for various naturally occurring alkaloid opioid ligands, and subsequently confirmed by molecular cloning. The amino acid sequences of the opioid receptor subtypes are ~70% homologous, and are similar to somatostatin receptors (SSTRs) showing ~40 % homology with SSTR1. G-protein binding is thought to occur at the third intracellular loop of the opioid receptors, which is also the location of consensus sequences for phosphorylation of the receptor. Interestingly, the genes encoding the specific receptor subtypes are found on different chromosomes in both the human and mouse genomes.
Immunohistochemistry analysis of paraffin-embedded human brain tissue using Delta Opioid Receptor (phospho S363) antibody (ab62152) at 1/50 - 1/100 dilution, in the presence (right panel) and absence (left panel) of immunising phosphopeptide.
ICC/IF image of ab62152 stained SKNSH cells. The cells were 4% formaldehyde (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab62152, 10µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899 Dylight 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
References for Anti-Delta Opioid Receptor (phospho S363) antibody (ab62152)
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