Akt Substrate, 160kD, phosphorylated (Thr642) (AS160, DKFZp779C0666, KIAA0603, TBC1 domain family member 4, Rab GAP)

AS160 is a 160kD Akt substrate that plays a critical role in regulating insulin-stimulated exocytosis of glucose transporter GLUT4. AS160 contains a Rab GTPase-activating protein domain (GAP), suggesting its potential role in membrane trafficking. Insulin-stimulated phosphorylation of AS160 on threonine 642 is critical for mediating GLUT4 translocation in fat and muscle cells, via inactivation of Rab GAP function. Phosphorylation of threonine 642 is also important in mediating AS160 redistribution from low density microsomes to the cytosol in adipocytes. \n\nApplications: \nSuitable for use in ELISA and Western Blot. \nOther applications not tested.\n\nRecommended Dilution:\nWestern Blot: 1:1000\nOptimal dilutions to be determined by the researcher.\n\nPositive Control: \n3T3-L1 adipocytes or COS cells overexpressing AS160, treated with insulin.\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. For long-term storage, store at -20 degrees C. Aliquots are stable for at least 12 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

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SPECIFICATIONS

Catalog Number

A1124-53

Size

10 Blots

Applications

ELISA, WB

Hosts

Rabbit

Reactivities

Hum

Form

Supplied as a liquid in PBS (without Mg2+ and Ca2+), 1mg/ml BSA (IgG, protease free), pH 7.2, 0.1% sodium azide, 50% glycerol.

P Type

Pab

Purity

Purified from rabbit serum by affinity chromatography.

References

1. Zeigerer, A. et al. (2004) Insulin stimulation of GLUT4 exocytosis, but not its inhibition of endocytosis, is dependent on RabGAP AS160. Mol. Biol. Cell. 15(10):4406-4415. 2. Sano, H. et al. (2003) Insulin-stimulated phosphorylation of a Rab GTPase-activating protein regulates GLUT4 translocation. J. Biol. Chem. 278(17):14599-14602. 3. Kane, S. et al. (2002) A method to identify serine kinase substrates. Akt phosphorylates a novel adipocyte protein with a Rab GTPase-activating protein (GAP) domain. J. Biol. Chem. 277(25):22115-22118. 4. Lavan, B.E. et al (1997) A novel 160-kDa phosphotyrosine protein in insulin-treated embryonic kidney cells is a new member of the insulin receptor substrate family. J. Biol. Chem 272(34):21403-21407.

Additional Info

Recognizes human AS160 when phosphorylated at Thr642. Species crossreactivity: Mouse and rat.