For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
NOTE: Please refer to primary antibody product webpage for recommended antibody dilution.
From sample preparation to detection, the reagents you need for your Western Blot are now in one convenient kit: #12957 Western Blotting Application Solutions Kit
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).
NOTE: Volumes are for 10 cm x 10 cm (100 cm2) of membrane; for different sized membranes, adjust volumes accordingly.
* Avoid repeated exposure to skin.
posted June 2005
revised June 2020
Protocol Id: 10
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Lys192 of human GPRIN1 protein.
G protein-regulated inducer of neurite outgrowth 1 (GPRIN1) and the two human homologs GPRIN2 and GPRIN3 are Gαz-binding proteins. The expression of these proteins and the activation of Gαo spurs neurite outgrowth in neurons. The function of these proteins include the G-protein-coupled receptors pathways, which catalyze the GDP-GTP exchange of heterodimeric G proteins, where the activation occurs when the GTP-bound form of G-protein α subunits, and in some cases βγ subunits are released (1).
GPRIN1 plays a role in the activation of nicotinic acetylcholine pathways by interacting with β2 subunits of nicotinic acetylcholine receptors (nAChRs) and other nAChR-interacting proteins, including G protein α and the G protein-activated K+ channel 1 (2). The observed neurite outgrowth of neurons in the CA3 and CA1 hippocampus is facilitated by interaction with the α7 subunit of nAChR, which is dependent on interaction with the G protein complex pathway (GPC), including Gαi/o (3,4).
Additionally, the interaction of GPRIN1 with the serotonin receptor 5-HT6 potentiates the 5-HT6R constitutive activation of the Gs-coupled receptors pathways, promoting neurite elongation and branching of striatal neurons. CDK5 interacts with the 5-HT6R C-terminal domain resulting in the phosphorylation of 5-HT6R at Ser350, increasing the affinity of GPRIN1 (5).
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