Cell Signaling Technology

Product Pathways - Tyrosine Kinase / Adaptors

Phospho-ALK (Tyr1604) Antibody #3341

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

Applications Key:  W=Western Blotting  IP=Immunoprecipitation
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 (Tyr1604) Antibody detects ALK only when phosphorylated at Tyr1604 (equivalent to Tyr664 of NPM-ALK). This antibody may cross-react with other activated protein tyrosine kinases including EGFR.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr1604 of human ALK. Antibodies are purified by protein A and peptide affinity chromatography

Western Blotting

Western Blotting

Western blot analysis of extracts from Sup-M2 cells, using Phospho-ALK (Tyr1604) 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. (Sup-M2 cells provided by Dr. Stephan W. Morris, St. Jude Children's Research Hospital, Tennessee.)

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 a nucleophosmin (NPM)-ALK fusion protein produced by a translocation (4). Investigators have found that the NPM-ALK fusion protein is a constitutively active, oncogenic tyrosine kinase associated with anaplastic lymphoma (4). Research literature suggests that activation of PLCγ by NPM-ALK may be a crucial step for its mitogenic activity and involved in the pathogenesis of anaplastic lymphomas (5).A distinct ALK oncogenic fusion protein involving ALK and echinoderm microtubule-associated protein like 4 (EML4) has been described in the research literature from a non-small cell lung cancer (NSCLC) cell line, with corresponding fusion transcripts 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 (6-8).

Phosphorylated Tyr664 of NPM-ALK (equivalent to Tyr1604 of full length ALK) is required for the interaction with PLCgamma (5). Site-directed mutagenesis of this tyrosine residue results in the loss of oncogenic activity of NPM-ALK (5).

  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.
  8. Soda, M. et al. (2007) Nature 448, 561-6.

Application References

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!

Companion Products

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

Products