Immunohistochemistry analysis of GPR32 was performed on neuroendocrine cells in human small intestine 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 GPR32 polyclonal antibody (Product # PA3-021) 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-LARAFGEEEFLSS-Abu-PRGNAPRE|
|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 GPR32 in human small intestine tissue neuroendocrine cells.
WB analysis shows GPR32 in glycoprotein-enriched fractions from GPR32 overexpressing 293 cells. Additional unknown bands at ~30kD and ~130kD were also detected.
While GPR32 has a theoretical MW of ~ 40kD, the observed band in a WB runs at ~60kD 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. The intronless GPR32 gene encodes an orphan G protein-coupled receptor that binds to resolvin D1 and lipoxin A4. Resolvin D1 binding to GPR32 has been linked to resolution of pulmonary inflammation, suppression of TGFbeta induced epithelial mesenchymal transition of A549 lung cancer cells. Resolvins are being evaluated as new drug candidates acting at the resolution phase of chronic inflammation.
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