Cell Signaling Technology

Product Pathways - PI3K / Akt Signaling

Phospho-NDRG1 (Thr346) Antibody #3217

Applications Reactivity Sensitivity MW (kDa) Source
W IP H M R (Mk) Endogenous 46, 48 Rabbit

Applications Key:  W=Western Blotting  IP=Immunoprecipitation
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey
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-NDRG1 (Thr346) Antibody detects endogenous levels of NDRG1 when phosphorylated at Thr346. This antibody likely cross-reacts with other conserved phosporylation sites on NDRG1 at positions Thr356 and Thr366.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr346 of NDRG1. Antibodies are purified by protein A and peptide affinity chromaography.

Western Blotting

Western Blotting

Western blot analysis of extracts from C2C12 cells, untreated, λ phosphatase-treated or treated with insulin for 30 minutes, using Phospho-NDRG1 (Thr346) Antibody (upper) or Actin Antibody #4967 (lower).

Background

N-myc downstream-regulated gene 1 (NDRG1), also termed Cap43, Drg1, RTP/rit42, and Proxy-1, is a member of the NDRG family, which is composed of four members (NDRG1-4) that function in growth, differentiation, and cell survival (1-5). NDRG1 is ubiquitously expressed and highly responsive to a variety of stress signals including DNA damage (4), hypoxia (5), and elevated levels of nickel and calcium (2). Expression of NDRG1 is elevated in N-myc defective mice and is negatively regulated by N- and c-myc (1,6). During DNA damage, NDRG1 is induced in a p53-dependent fashion and is necessary for p53-mediated apoptosis (4,7). Research studies have shown that NDRG1 may also play a role in cancer progression by promoting differentiation, inhibiting growth, and modulating metastasis and angiogenesis (3,4,6,8,9). Nonsense mutation of the NDRG1 gene has been shown to cause hereditary motor and sensory neuropathy-Lom (HMSNL), which is supported by studies demonstrating the role of NDRG1 in maintaining myelin sheaths and axonal survival (10,11). NDRG1 is up-regulated during mast cell maturation and its deletion leads to attenuated allergic responses (12). Both NDRG1 and NDRG2 are substrates of SGK1, although the precise physiological role of SGK1-mediated phosphorylation is not known (13). NDRG1 is phosphorylated by SGK1 at Thr328, Ser330, Thr346, Thr356, and Thr366. Phosphorylation by SGK1 primes NDRG1 for phosphorylation by GSK-3.

Phospho-NDRG1 (Thr346) Antibody is directed at a site that was identified at Cell Signaling Technology (CST) using PhosphoScan®, CST's LC-MS/MS platform for modification site discovery. Phosphorylation at Thr346 was discovered using an Akt substrate antibody and was shown to be induced by insulin treatment in multiple cell lines. Please visit PhosphoSitePlus®, CST's modification site knowledgebase, at www.phosphosite.org for more information.

  1. Shimono, A. et al. (1999) Mech Dev 83, 39-52.
  2. Zhou, D. et al. (1998) Cancer Res 58, 2182-9.
  3. van Belzen, N. et al. (1997) Lab Invest 77, 85-92.
  4. Kurdistani, S.K. et al. (1998) Cancer Res 58, 4439-44.
  5. Park, H. et al. (2000) Biochem Biophys Res Commun 276, 321-8.
  6. Li, J. and Kretzner, L. (2003) Mol Cell Biochem 250, 91-105.
  7. Stein, S. et al. (2004) J Biol Chem 279, 48930-40.
  8. Maruyama, Y. et al. (2006) Cancer Res 66, 6233-42.
  9. Nishio, S. et al. (2008) Cancer Lett 264, 36-43.
  10. Kalaydjieva, L. et al. (2000) Am J Hum Genet 67, 47-58.
  11. Okuda, T. et al. (2004) Mol Cell Biol 24, 3949-56.
  12. Taketomi, Y. et al. (2007) J Immunol 178, 7042-53.
  13. Murray, J.T. et al. (2004) Biochem J 384, 477-88.

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