Immunohistochemistry analysis of GPR39 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 GPR39 polyclonal antibody (Product # PA3-027) 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-NSGAKPANSAAENGFQEHEV|
|Storage buffer||whole serum|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|Immunohistochemistry (Paraffin) (IHC (P))||1:3000|
|Western Blot (WB)||1:1000-1:5000|
* 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 GPR39 in human enteric ganglion.
WB analysis shows GPR39 in glycoprotein-enriched fractions from mock transfected or GPR39 overexpressing 293 cells. The GPR39 antiserum showed specific detection of GPR39 over other tested GPRs.
While GPR39 has a theoretical MW of ~ 51kD, the observed band in a WB runs at ~72kD 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. G protein-coupled receptor 39 is an orphan receptor in the Ghrelin family pathway. Various studies have shown that GPR39 plays important roles in hypertension, adipogenesis and that overexpression of GPR39 contributes to malignant development of human esophageal squamous cell carcinoma.
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