Immunofluorescence analysis of G Protein alpha Inhibitor 1 and 2 was performed using 70% confluent log phase SH-SY5Y cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with GNAI1/GNAI2 Rabbit Polyclonal Antibody (PA1-1000) at 2ug/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic and membranous localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse, Rat|
|Published species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues K(346) N N L K D C G L F(355) of human G Protein alpha Inhibitor 1/2.|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Immunoprecipitation (IP)||Assay dependent|
|Western Blot (WB)||1 µg/ml|
* 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.
|Immunoprecipitation (IP)||See 1 publications below|
PA1-1000 detects G protein alpha inhibitor 1/2 from human, rat, and mouse tissues and cells.
PA1-1000 has been successfully used in Immunoprecipitation and Western blot procedures. By Western blot, this antibody detects an ~38 kDa protein representing G protein alpha inhibitor 1/2 from HeLa cell lysate.
The PA1-1000 immunogen is a synthetic peptide corresponding to residues K(346) N N L K D C G L F(355) of human G Protein alpha Inhibitor 1/2. This sequence is completely conserved in human, rat, guinea pig, hamster, bovine, hydra, octopus, Xenopus, chicken, canine, spiny dogfish, great pond snail, starfish, and American lobster G protein alpha inhibitor 1/2. PA1-1000 immunizing peptide (Cat. # PEP-129) is available for use in neutralization and control procedures.
The G protein family of signal transducers includes the alpha, beta, gamma subunits and the effector which are most clearly distinguished by their different alpha chains. The family also includes Gs and Gi, the stimulatory and inhibitory GTP-binding regulators of adenylate cyclase; Go, a protein of unknown function that is abundantly expressed in brain; and transducin 1 and 2, proteins involved in retinal phototransduction. The G proteins possess GTPase activity and are components of a complex membrane signaling system that includes membrane-bound receptors. The system exists to modulate extracellular signals as they are transmitted into the cell.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Gut-expressed gustducin and taste receptors regulate secretion of glucagon-like peptide-1.
PA1-1000 was used in immunoprecipitation to investigate the effects of gustducin and taste receptors on glucagon-like peptide-1
|Jang HJ,Kokrashvili Z,Theodorakis MJ,Carlson OD,Kim BJ,Zhou J,Kim HH,Xu X,Chan SL,Juhaszova M,Bernier M,Mosinger B,Margolskee RF,Egan JM||Proceedings of the National Academy of Sciences of the United States of America (104:15069)||2007|