AS160 Signaling Antibody Sampler Kit #59633

Kit Usage Information

Protocols

• 2670: Western Blotting, Immunoprecipitation (Magnetic)
• 4060: Western Blotting, Immunoprecipitation (Agarose), Immunohistochemistry (Paraffin), Immunofluorescence, Flow
• 4691: Western Blotting, Immunoprecipitation (Agarose), Immunohistochemistry (Paraffin), Immunofluorescence, Flow
• 7074: Western Blotting
• 8619: Western Blotting, Immunoprecipitation (Magnetic)
• 8730: Western Blotting, Immunoprecipitation (Magnetic)
• 8881: Western Blotting
• 13038: Western Blotting, Immunoprecipitation (Magnetic), Immunofluorescence, Flow

Product Description

The AS160 Signaling Antibody Sampler Kit provides an economical means of detecting select components involved in the AS160 signaling pathway. The kit contains enough primary antibodies to perform at least two western blot experiments per antibody.

Specificity / Sensitivity

AS160 (C69A7) Rabbit mAb detects endogenous levels of total AS160 protein. Phospho-AS160 (Ser318) (D3D11) Rabbit mAb recognizes endogenous levels of AS160 protein only when phosphorylated at Ser318. Phospho-AS160 (Ser588) (D8E4) Rabbit mAb recognizes endogenous levels of AS160 protein only when phosphorylated at Ser588. Phospho-AS160 (Thr642) (D27E6) Rabbit mAb recognizes endogenous levels of AS160 protein only when phosphorylated at Thr642. Akt (pan) (C67E7) Rabbit mAb detects endogenous levels of total Akt protein. This antibody does not cross-react with other related proteins. Phospho-Akt (Thr308) (D25E6) XP^® Rabbit mAb recognizes endogenous levels of Akt1 protein only when phosphorylated at Thr308. This antibody also recognizes endogenous levels of Akt2 protein when phosphorylated at Thr309 or Akt3 protein when phosphorylated at Thr305. Phospho-Akt (Ser473) (D9E) XP^® Rabbit mAb detects endogenous levels of Akt only when phosphorylated at Ser473.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ala195 of human AS160. Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser318 of human AS160 protein. Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser588 of human AS160 protein. Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr642 of human AS160 protein. Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues in the carboxy-terminal sequence of mouse Akt. Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Thr308 of human Akt1 protein. Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues around Ser473 of human Akt.

Background

Insulin is a major hormone controlling critical energy functions, such as glucose and lipid metabolism. Insulin binds to and activates the insulin receptor (IR) tyrosine kinase, which phosphorylates and recruits adaptor proteins. The signaling pathway initiated by insulin and its receptor stimulates glucose uptake in muscle cells and adipocytes through translocation of the Glut4 glucose transporter from the cytoplasm to the plasma membrane (1). A 160 kDa substrate of the Akt Ser/Thr kinase (AS160, TBC1D4) is a Rab GTPase-activating protein that regulates insulin-stimulated Glut4 trafficking. AS160 is expressed in many tissues including brain, kidney, liver, and brown and white fat (2). Multiple Akt phosphorylation sites have been identified on AS160 in vivo, with five sites (Ser318, Ser570, Ser588, Thr642, and Thr751) showing increased phosphorylation following insulin treatment (2,3). Studies using recombinant AS160 demonstrate that insulin-stimulated phosphorylation of AS160 is a crucial step in Glut4 translocation (3) and is reduced in some patients with type 2 diabetes (4). The interaction of 14-3-3 regulatory proteins with AS160 phosphorylated at Thr642 is a necessary step for Glut4 translocation (5). Phosphorylation of AS160 by AMPK is involved in the regulation of contraction-stimulated Glut4 translocation (6).
Akt, also referred to as PKB or Rac, plays a critical role in controlling survival and apoptosis (7-9). This protein kinase is activated by insulin and various growth and survival factors to function in a wortmannin-sensitive pathway involving PI3 kinase (8,9). Akt is activated by phospholipid binding and activation loop phosphorylation at Thr308 by PDK1 (10) and by phosphorylation within the carboxy terminus at Ser473. The previously elusive PDK2 responsible for phosphorylation of Akt at Ser473 has been identified as the mammalian target of rapamycin (mTOR) in a rapamycin-insensitive complex with rictor and Sin1 (11,12).

1. Watson, R.T. and Pessin, J.E. (2006) Trends Biochem. Sci. 31, 215-22.
2. Kane, S. et al. (2002) J. Biol. Chem. 277, 22115-8.
3. Sano, H. et al. (2003) J. Biol. Chem. 278, 14599-602.
4. Karlsson, H.K. et al. (2005) Diabetes 54, 1692-7.
5. Ramm, G. et al. (2006) J. Biol. Chem. 281, 29174-80.
6. Kramer, H.F. et al. (2006) J. Biol. Chem. 281, 31478-85.
7. Franke, T.F. et al. (1997) Cell 88, 435-7.
8. Burgering, B.M. and Coffer, P.J. (1995) Nature 376, 599-602.
9. Franke, T.F. et al. (1995) Cell 81, 727-36.
10. Alessi, D.R. et al. (1996) EMBO J 15, 6541-51.
11. Sarbassov, D.D. et al. (2005) Science 307, 1098-101.
12. Jacinto, E. et al. (2006) Cell 127, 125-37.

Pathways & Proteins

Explore pathways + proteins related to this product.

Select Your Pathway/Protein AMPK Signaling Adherens Junction Dynamics Alzheimer's Disease Angiogenesis Apoptosis Regulation B Cell Receptor Signaling ESC Pluripotency and Differentiation ErbB/HER Signaling G1/S Checkpoint IL6 Signaling Inhibition of Apoptosis Insulin Receptor Signaling Microtubule Dynamics Mitochondrial Control of Apoptosis NF-kB Signaling PI3K/Akt Signaling Protein Kinase C Signaling T Cell Receptor Signaling TGF-ß Signaling Translation: eIF4E and p70S6K Warburg Effect mTOR Signaling Protein: O60343 Protein: P31749 Protein: P31751 Protein: Q9Y243

Database Links

UniProt ID: O60343 , P31751 , Q9Y243 , P31749

Entrez-Gene Id: 9882 , 208 , 10000 , 207

View in PhosphoSitePlus^®