Cell Signaling Technology

Product Pathways - Metabolism

Phospho-TBC1D1 (Ser660) Antibody #6928

Applications Reactivity Sensitivity MW (kDa) Source
W M (R) Transfected Only 160 Rabbit

Applications Key:  W=Western Blotting
Reactivity Key:  M=Mouse  R=Rat
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

Phospho-TBC1D1 (Ser660) Antibody recognizes transfected levels of TBC1D1 protein only when phosphorylated at Ser660.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser660 of mouse TBC1D1 protein. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293T cells, transfected with either mouse wild-type TBC1D1 or mutant TBC1D1 (S660A), using Phospho-TBC1D1 (Ser660) Antibody (upper) or TBC1D1 (G689) Antibody #5929 (lower). Both expression vectors were kindly provided by Dr. Laurie Goodyear at the Joslin Diabetes Center.

Background

TBC1D1 is a paralog of AS160 (1) and both proteins share about 50% identity (2). TBC1D1 was shown to be a candidate gene for severe obesity (3). It plays a role in Glut4 translocation through its GAP activity (2,4). Studies indicate that TBC1D1 is highly expressed in skeletal muscle (1). Insulin, AICAR, and contraction directly regulate TBC1D1 phosphorylation in this tissue (1). Three AMPK phosphorylation sites (Ser231, Ser660, and Ser700) and one Akt phosphorylation site (Thr590) were identified (5). Muscle contraction or AICAR treatment increases phosphorylation on Ser231, Ser660, and Ser700 but not on Thr590; insulin increases phosphorylation on Thr590 only (5).

  1. Taylor, E.B. et al. (2008) J Biol Chem 283, 9787-96.
  2. Roach, W.G. et al. (2007) Biochem J 403, 353-8.
  3. Stone, S. et al. (2006) Hum Mol Genet 15, 2709-20.
  4. Chavez, J.A. et al. (2008) J Biol Chem 283, 9187-95.
  5. Vichaiwong, K. et al. (2010) Biochem J 431, 311-20.

Application References

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Companion Products

For Research Use Only. Not For Use In Diagnostic Procedures.

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