Immunofluorescence analysis of Galphai1 (GNAI1) was performed using 70% confluent log phase RSC-96 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 (R4.5) Mouse Monoclonal Antibody (MA5-12800) at 2ug/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (A28175) 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 membranous localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Bovine, Guinea pig, Human, Mouse, Rat|
|Published species reactivity||Rat, Human|
|Host / Isotype||Mouse / IgG2b|
|Immunogen||Partially purified rat brain Galphai1 protein|
|Storage buffer||PBS, pH 7.4, with 0.2% BSA|
|Contains||0.09% sodium azide|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Western Blot (WB)||1-3 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.
|Western Blot (WB)||See 2 publications below|
MA5-12800 targets Galphai1 G-Protein in WB applications and shows reactivity with Bovine, Guinea Pig, Human, mouse, and Rat samples.
The MA5-12800 immunogen is partially purified rat brain Galphai1 protein.
GTP-binding proteins (G-proteins) play a critical role in signal transduction by coupling receptors to effectors. The intracellular consequences of G-protein activation include second messenger generation, protein phosphorylation, ion channel activation, gene induction, cell growth, and differentiation. Gi belongs to the pertussis toxin sensitive G-proteins. Gi consists of three different subtypes of alpha-subunits (alphai1, alphai2 and alphai3)
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Human neuroglobin functions as an oxidative stress-responsive sensor for neuroprotection.
MA5-12800 was used in western blot to study the role of neuroglobin as an oxidative stress sensor promoting neuronal survival under conditions of oxidative stress
|Watanabe S,Takahashi N,Uchida H,Wakasugi K||The Journal of biological chemistry (287:30128)||2012|
Molecular basis of guanine nucleotide dissociation inhibitor activity of human neuroglobin by chemical cross-linking and mass spectrometry.
MA5-12800 was used in western blot to investigate the details of neuroglobin and Galpha(i) interaction
|Kitatsuji C,Kurogochi M,Nishimura S,Ishimori K,Wakasugi K||Journal of molecular biology (368:150)||2007|