Immunohistochemistry analysis of GPR18 was performed on human enteric ganglion tissue. To expose target proteins, antigen retrieval was performed by microwaving tissues for 20 minutes in 10mM sodium citrate buffer (pH 6.0). Tissue slides were probed with a GPR18 polyclonal antibody (Product # PA3-006) at a dilution of 1:3000, overnight at 4C in a humidified chamber. Tissues were washed, and detection was performed using an ABC kit composed of biotinylated goat anti-rabbit IgG, peroxidase-conjugated avidin, and 3-amino-9-ethylcarbazole (AEC) substrate in acetate buffer. Tissues were counterstained with hematoxylin and dehydrated to prep for mounting.
|Tested species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||KLH conjugated Cys-RKSFRSGSLRSLSNINSEML|
|Storage buffer||whole serum|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|Immunohistochemistry (Paraffin) (IHC (P))||1:3000|
|Western Blot (WB)||1:1000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
IHC (P) analysis shows positive staining of GPR18 in human enteric ganglion tissue.
WB analysis shows GPR18 in glycoprotein-enriched fractions from GPR18 overexpressing 293 cells. A band of similar MW was also detected in GPR3 overexpressing 293 cells, suggesting some cross-reactivity with GPR3 or enhanced detection of basal GPR18 in the enriched fractions.
While GPR18 has a theoretical MW of ~ 38kD, the observed band in a WB runs at ~70kD due to enrichment of the glycosylated receptor.
The G-protein-coupled receptor (GPCR) superfamily is comprised of an estimated 600-1,000 members and is the largest known class of molecular targets with proven therapeutic value. GPR18 is an orphan G protein-coupled receptor found at high levels in the testis, small intestines and cells associated with the immune system including lymphocytes, thymus and spleen. GPR18 has been found to be a receptor for endogenous lipid neurotransmitters, several of which also bind to cannabinoid receptors. Some ligands described to be GPR18 agonists or partial agonists include N-Arachidonoylglycine, Abnormal cannabidiol, O-1602, AM-251, Cannabidiol, delta 9-Tetrahydrocannabinol, N-arachidonoyl ethanolamine and Arachidonylcyclopropylamide. Antagonists include the synthetic cannabidiol related compounds CID-85469571 and O-1918.
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