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Western blot analysis of GIT1 on a rat cerebrum lysate. Lane 1: 1:250, lane 2: 1:500, lane 3: 1: 1000 dilution of the mouse anti-GIT1 antibody.
Immunofluorescence staining of A431cells (Human epithelial carcinoma; ATCC CRL-1555) with the mouse anti-GIT1 antibody.
BD Transduction Laboratories™ Purified Mouse Anti-GIT1
BD Transduction Laboratories™ Purified Mouse Anti-GIT1
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准备和存储
推荐的实验流程
Western blot: Please refer to http://www.bdbiosciences.com/pharmingen/protocols/Western_Blotting.shtml
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- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
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Activation of adrenergic receptors (AR), such as β2AR leads to activation of heterotrimeric G proteins and recruitment of G protein-coupled receptor kinases (GRKs). Phosphorylation of the activated receptor by GRKs promotes the binding of arrestin protein which prevents further G protein activation. GRK interactor 1 (GIT1) was identified in a yeast two-hybrid screen for proteins that bind GRK2. GIT1 has an N-terminal Zn2+ finger-like motif involved in GTPase activating activity and multipe ankyrin repeat units. In vitro, GIT1 is a GTPase-activating protein (GAP) for the ADP ribosylation factor (ARF) family of small GTP-binding proteins. In rat, GIT1 is widely expressed with the highest levels in the testis and lowest levels in the liver and spleen. Overexpression of GIT1 in HEK293 cells causes reduced β2AR signaling and increased receptor phosphorylation as a result of reduced receptor internalization and resensitization. In addition, GIT1 interacts directly with paxillin and the PIX exchange factors, and indirectly with PAKs, Rac1, and Nck. GIT proteins are also now known to be PIP3-stimulated GAPs for ARF6. Thus, GIT1 activation of ARF proteins may involve cell signaling pathways that are important for endocytosis, cell adhesion, and cytoskeletal dynamics.
研发参考 (2)
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Premont RT, Claing A, Vitale N. beta2-Adrenergic receptor regulation by GIT1, a G protein-coupled receptor kinase-associated ADP ribosylation factor GTPase-activating protein. Proc Natl Acad Sci U S A. 1998; 95(24):14082-14087. (Biology). 查看参考
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Zhao ZS, Manser E, Loo TH, Lim L. Coupling of PAK-interacting exchange factor PIX to GIT1 promotes focal complex disassembly. Mol Cell Biol. 2000 ; 20(17):6354-6363. (Biology). 查看参考
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