N-arachidonyl glycine receptor also known as G-protein coupled receptor 18 (GPR18) is a protein that in humans is encoded by the GPR18 gene. Along with the other previously "orphan" receptors GPR55 and GPR119, GPR18 has been found to be a receptor for endogenous lipid neurotransmitters, several of which also bind to cannabinoid receptors. Research supports the hypothesis that GPR18 is the abnormal cannabidiol receptor and N-arachidonoyl glycine, the endogenous lipid metabolite of anandamide, initiates directed microglial migration in the CNS through activation of GPR18. Anti-GPR18 - http://www.stjohnslabs.com/gpr18-antibody-p-96212?filter_name=STJ93370 Join our Antibody Validation Project - http://www.stjohnslabs.com/services/antibody-validation

1. Result
“Good; several bands of varying molecular weights appear, but the predicted 34 kDa band is evident .” - A. Cassey, Ex-
ternal Laboratory.
Cell Line:
Whole cell lysates from 12 hu-
man vulvar fibroblast strains
Method of validation: Western Blot
Primary Antibody:
STJ93370 Anti-GPR18 Anti-
body
Secondary Antibody HRP goat anti-rabbit
Dilution ratio: 1:2000
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STJ93370 Anti-GPR18 Antibody
Antibody Specificity:
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Protocol
Treatment of materials: whole cell lysates from 12 human vulvar fibroblast strains
E.g. Gel electrophoresis: SDS-polyacrylamide gel with a 5% acrylamide stacking gel
Transfer: Electrophoresed proteins were electro-transferred to PVDF membrane (Millipore) overnight at 25 V in a
standard TRIS-Glycine-Methanol transfer buffer
Blocking: The Western blots were then blocked with 2% BSA in PBS for 60 min. at 4*C.
Primary antibody probing: primary antibody was added to 2% BSA in PBS + 0.1% Tween-20 for 12 hrs. at 4*C.
Membrane wash: The Western blots were washed with moderate agitation 5 times for 5 min. each in PBS + 0.1%
Tween-20.
Secondary antibody probing: at 1:20,000 dilution in 2% BSA in PBS + 0.1% Tween-20 for 2 hrs. at 4*C.
Membrane wash: The Western blots were washed with moderate agitation 5 times for 5 min. each in PBS + 0.1%
Tween-20.
Visualization: ECL visualization for 5 min (the visualization time was determined by actual result. It was ensured that
the visualization time was identical for the same antigen).
Figure: Western blot of whole cell lysates from 12
human vulvar fibroblast strains
8 ug of total protein per gel lane (far left and right
lanes contained Mol. Wt. markers)