Cell Signaling Technology

Product Pathways - Tyrosine Kinase / Adaptors

Phospho-ALK (Tyr1586) Antibody #3343

Applications Reactivity Sensitivity MW (kDa) Source
W H Endogenous 80 (NPM-ALK) 220 (ALK) Rabbit

Applications Key:  W=Western Blotting
Reactivity Key:  H=Human
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-ALK (Tyr1586) Antibody detects ALK only when phosphorylated at tyrosine 1586 (equivalent to Tyr646 of NPM-ALK).

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr1586 of human ALK. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cells expressing different activated tyrosine kinases, using Phospho-ALK (Tyr1586) Antibody (upper) or Phospho-Tyrosine mAb (P-Tyr-100) #9411 (lower). Phospho-ALK (Tyr1586) Antibody shows no cross-reactivity with other tyrosine-phosphorylated tyrosine kinases.

Western Blotting

Western Blotting

Western blot analysis of extracts from Sup-M2 cells, using Phospho-ALK (Tyr1586) Antibody (A,B) or ALK Antibody (C,D). The phospho-specificity of the antibody was characterized by treating the membrane with calf intestinal alkaline phosphatase (CIP) (B,D) after western transfer.

Background

Anaplastic lymphoma kinase (ALK) is a tyrosine kinase receptor for pleiotrophin (PTN), a growth factor involved in embryonic brain development (1-3). In ALK-expressing cells, PTN induces phosphorylation of both ALK and the downstream effectors IRS-1, Shc, PLCγ, and PI3 kinase (1). ALK was originally discovered as an NPM (nucleophosmin)-ALK fusion protein produced by a translocation (4). The NPM-ALK fusion protein is a constitutively active, oncogenic tyrosine kinase associated with anaplastic lymphoma (4). Activation of PLCγ by NPM-ALK has been suggested to be a crucial step for its mitogenic activity and may be important in the pathogenesis of anaplastic lymphomas (5). A distinct ALK oncogenic fusion protein involving ALK and EML4 has been described from a non-small cell lung cancer cell line and corresponding fusion transcripts are present in some cases of lung adenocarcinoma. The short, amino-terminal region of the microtubule-associated protein EML4 is fused to the kinase domain of ALK (6,7).

  1. Stoica, G.E. et al. (2001) J Biol Chem 276, 16772-9.
  2. Iwahara, T. et al. (1997) Oncogene 14, 439-49.
  3. Morris, S.W. et al. (1997) Oncogene 14, 2175-88.
  4. Morris, S.W. et al. (1994) Science 263, 1281-4.
  5. Bai, R.Y. et al. (1998) Mol Cell Biol 18, 6951-61.
  6. Rikova, K. et al. (2007) Cell 131, 1190-203.
  7. Takeuchi, K. et al. (2008) Clin Cancer Res 14, 6618-24.

Application References

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Companion Products

This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

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