Cell Signaling Technology

Product Pathways - PI3K / Akt Signaling

Phospho-TSC2 Antibody Sampler Kit #8350

Kit Includes Quantity Applications Reactivity MW (kDa) Isotype
Tuberin/TSC2 (D93F12) XP® Rabbit mAb #4308 40 µl W IP IF-IC H M R Hm Mk 200 Rabbit IgG
Phospho-Tuberin/TSC2 (Ser939) Antibody #3615 40 µl W H M 200 Rabbit
Phospho-Tuberin/TSC2 (Thr1462) (5B12) Rabbit mAb #3617 40 µl W IP H M R 200 Rabbit IgG
Phospho-Tuberin/TSC2 (Ser1387) Antibody #5584 40 µl W IP H M R Mk 200 Rabbit
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl Goat

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:  H=Human  M=Mouse  R=Rat  Hm=Hamster  Mk=Monkey
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Specificity / Sensitivity

Each antibody in the Phospho-TSC2 Antibody Sampler Kit recognizes only its specific target and does not cross-react with other family members unless otherwise indicated. Phospho-Tuberin/TSC2 (Ser1387) Antibody detects endogenous levels of tuberin protein only when phosphorylated at Ser1387. This antibody also cross-reacts with an unidentified 140 kDa protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from NIH/3T3 cells, untreated, PDGF-treated, and PDGF and wortmannin-treated or PDGF and rapamycin-treated, using Phospho-Tuberin/TSC2 (Ser939) Antibody #3615 (top), Phospho-Tuberin/TSC2 (Thr1462) Antibody #3611 (middle) or Tuberin/TSC2 Antibody #3612 (bottom).

Western Blotting

Western Blotting

Western blot analysis of extracts from NIH/3T3 cells, treated with λ phosphatase or PDGF, using Phospho-Tuberin/TSC2 (Thr1462) (5B12) Rabbit mAb #3617 (upper) or Tuberin Antibody (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from wild-type (wt) and TSC2-deficient (-/-) mouse embryonic fibroblasts (MEF) using Tuberin/TSC2 (D93F12) XP® Rabbit mAb #4308 (upper) and Akt (pan) (C67E7) Rabbit mAb #4691 (lower).


Western Blotting

Western Blotting

Western blot analysis of extracts from SH-SY5Y cells, untreated or treated with AICAR #9944 (2 mM, 30 min), using Phospho-Tuberin/TSC2 (Ser1387) Antibody #5584 (upper) and Tuberin/TSC2 (D93F12) XP® Rabbit mAb #4308 (lower).

Description

The Phospho-TSC2 Antibody Sampler Kit provides an economical means to investigate protein folding within the cell. The kit contains enough primary and secondary antibodies to perform four western blot experiments per primary antibody.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to carboxy-terminal residues of human tuberin protein or a synthetic phosphopeptide corresponding to residues surrounding Thr1462 of human tuberin protein. Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues around Ser939 or corresponding to residues surrounding Ser1387 of human tuberin protein. Polyclonal antibodies are purified by protein A and peptide affinity chromatography.

Background

Tuberin is a product of the TSC2 tumor suppressor gene and an important regulator of cell proliferation and tumor development (1). Mutations in either TSC2 or the related TSC1 (hamartin) gene cause tuberous sclerosis complex (TSC), an autosomal dominant disorder characterized by development of multiple, widespread non-malignant tumors (2). Tuberin is directly phosphorylated at Thr1462 by Akt/PKB (3). Phosphorylation at Thr1462 and Tyr1571 regulates tuberin-hamartin complexes and tuberin activity (3-5). In addition, tuberin inhibits the mammalian target of rapamycin (mTOR), which promotes inhibition of p70 S6 kinase, activation of eukaryotic initiation factor 4E binding protein 1 (4E-BP1, an inhibitor of translation initiation), and eventual inhibition of translation (3,6,7).

  1. Soucek, T. et al. (1998) Proc. Natl. Acad. Sci. USA 95, 15653-15658.
  2. Sparagana, S.P. and Roach, E.S. (2000) Curr. Opin. Neurol. 13, 115-119.
  3. Manning, B. D. et al. (2002) Mol. Cell 10, 151-161.
  4. Aicher, L. D. et al. (2001) J. Biol. Chem. 276, 21017-21021.
  5. Dan, H. C. et al. (2002) J. Biol. Chem. 277, 35364-35370.
  6. Goncharova, E.A. et al. (2002) J. Biol. Chem. 277, 30958-30967.
  7. Inoki, K. et al. (2002) Nat. Cell Biol. 4, 648-657.

Application References

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Protocols

* Product-specific protocol.

Companion Products

Selected rabbit monoclonal antibodies are produced under license (granting certain rights including those under U. S. Patents No. 5,675,063 and in some instances 7,429,487) from Epitomics, Inc. 

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

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