Revision 1

#8350Store at -20C

1 Kit

(4 x 20 microliters)

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Mol. Wt Isotype/Source
Tuberin/TSC2 (D93F12) XP® Rabbit mAb 4308 20 µl 200 kDa Rabbit IgG
Phospho-Tuberin/TSC2 (Ser939) Antibody 3615 20 µl 200 kDa Rabbit 
Phospho-Tuberin/TSC2 (Thr1462) (5B12) Rabbit mAb 3617 20 µl 200 kDa Rabbit IgG
Phospho-Tuberin/TSC2 (Ser1387) Antibody 5584 20 µl 200 kDa Rabbit 
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

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 two western blot experiments per primary antibody.

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

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 U S A 95, 15653-8.
  2. Sparagana, S.P. and Roach, E.S. (2000) Curr Opin Neurol 13, 115-9.
  3. Manning, B.D. et al. (2002) Mol Cell 10, 151-62.
  4. Aicher, L.D. et al. (2001) J Biol Chem 276, 21017-21.
  5. Dan, H.C. et al. (2002) J Biol Chem 277, 35364-70.
  6. Goncharova, E.A. et al. (2002) J Biol Chem 277, 30958-67.
  7. Inoki, K. et al. (2002) Nat Cell Biol 4, 648-57.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

    Limited Uses

    Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

    Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

    Revision 1
    #8350

    Phospho-TSC2 Antibody Sampler Kit

    Phospho-TSC2 Antibody Sampler Kit: Image 1 Expand Image
    Simple Western™ analysis of lysates (0.1 mg/mL) from COS-7 cells using Tuberin/TSC2 (D93F12) XP® Rabbit mAb #4308. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 66-440 kDa separation module.
    Phospho-TSC2 Antibody Sampler Kit: Image 2 Expand Image
    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 (top), Phospho-Tuberin/TSC2 (Thr1462) Antibody #3611 (middle) or Tuberin/TSC2 Antibody #3612 (bottom).
    Phospho-TSC2 Antibody Sampler Kit: Image 3 Expand Image
    Western blot analysis of extracts from l phosphatase-treated or PDGF treated NIH/3T3 cells, using Phospho- Tuberin/TSC2 (Thr1462) (5B12) Rabbit mAb (upper) or Tuberin Antibody (lower).
    Phospho-TSC2 Antibody Sampler Kit: Image 4 Expand Image
    Western blot analysis of extracts from SH-SY5Y cells and rat brain using Tuberin/TSC2 (D93F12) XP® Rabbit mAb.
    Phospho-TSC2 Antibody Sampler Kit: Image 5 Expand Image
    Western blot analysis of extracts from SH-SY5Y cells, untreated or treated with AICAR #9944 (2 mM for 30 minutes), using Phospho-Tuberin/TSC2 (Ser1387) Antibody (upper) and Tuberin/TSC2 (D93F12) XP® Rabbit mAb #4308 (lower).
    Phospho-TSC2 Antibody Sampler Kit: Image 6 Expand Image
    After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
    Phospho-TSC2 Antibody Sampler Kit: Image 7 Expand Image
    Western blot analysis of extracts from wild-type (wt) and TSC2-deficient (-/-) mouse embryonic fibroblasts (MEF) using Tuberin/TSC2 (D93F12) XP® Rabbit mAb (upper) and Akt (pan) (C67E7) Rabbit mAb #4691 (lower).
    Phospho-TSC2 Antibody Sampler Kit: Image 8 Expand Image
    Western blot analysis of extracts from SH-SY5Y cells, treated with AICAR #9944 (2 mM for 30 minutes), using Phospho-Tuberin/TSC2 (Ser1387) Antibody. The phospho-specificity of the antibody was verified by treating the membrane with (+) or without (-) calf intestinal phosphatase (CIP) after western transfer.
    Phospho-TSC2 Antibody Sampler Kit: Image 9 Expand Image
    Western blot analysis of extracts from HAP1 WT (left) or TSC2 KO (right) using Tuberin/TSC2 (D93F12) XP® Rabbit mAb (upper). Membranes stained with Ponceau S for total protein normalization (lower). These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibodies, as a companion to validation data generated by CST scientists.
    Phospho-TSC2 Antibody Sampler Kit: Image 10 Expand Image
    Confocal immunofluorescent analysis of SH-SY5Y (left), wild-type MEF (middle) and TSC2-deficient (-/-) MEF cells (right) using Tuberin/TSC2 (D93F12) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    Phospho-TSC2 Antibody Sampler Kit: Image 11 Expand Image
    Flow cytometric analysis of wild-type MEF (green) and TSC2-deficient (-/-) MEF cells (blue) using Tuberin/TSC2 (D93F12) XP® Rabbit mAb. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.