Immunohistochemistry analysis of NOP 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 NOP polyclonal antibody (Product # PA3-029) 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-VRSIAKDVALACKTSETVPRPA|
|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 NOP in human enteric ganglion.
WB analysis shows NOP in glycoprotein-enriched fractions from mock transfected or NOP overexpressing 293 cells.
While NOP has a theoretical MW of ~ 41kD, 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. The nociceptin receptor is a G-protein coupled opioid receptor that functions as receptor for the endogenous neuropeptide nociceptin. Ligand binding causes a conformation change that triggers signaling via guanine nucleotide-binding proteins (G proteins) and modulates the activity of down-stream effectors. Signaling via G proteins mediates inhibition of adenylate cyclase activity and calcium channel activity. Arrestins modulate signaling via G proteins and mediate the activation of alternative signaling pathways that lead to the activation of MAP kinases. Plays a role in modulating nociception and the perception of pain. Plays a role in the regulation of locomotor activity by the neuropeptide nociceptin.
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