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.
Western blot analysis of extracts from SH-SY5Y cells and rat brain using Tuberin/TSC2 (D93F12) XP® Rabbit mAb.
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).
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).
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).
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).
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).
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.
|Tuberin/TSC2 (D93F12) XP® Rabbit mAb 4308||20 µl||
||H M R Hm Mk||200||Rabbit IgG|
|Phospho-Tuberin/TSC2 (Ser939) Antibody 3615||20 µl||
|Phospho-Tuberin/TSC2 (Thr1462) (5B12) Rabbit mAb 3617||20 µl||
||H M R||200||Rabbit IgG|
|Phospho-Tuberin/TSC2 (Ser1387) Antibody 5584||20 µl||
||H M R Mk||200||Rabbit|
|Anti-rabbit IgG, HRP-linked Antibody 7074||100 µl||
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.
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.
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.
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).
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