Cell Signaling Technology

Product Pathways - Tyrosine Kinase / Adaptors

Phospho-ALK (Tyr1278) (D59G10) Rabbit mAb #6941

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

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 (Tyr1278) (D59G10) Rabbit mAb detects endogenous levels of ALK only when phosphorylated at Tyr1278, which is equivalent to Tyr338 of NPM-ALK. This antibody also reacts with leukocyte tyrosine kinase (LTK) phosphorylated at Tyr672, and may cross-react with other activated protein tyrosine kinases such as EGFR.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr1278 of human ALK protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from SUP-M2 cells, untreated or treated with calf intestinal phosphatase (CIP), using Phospho-ALK (Tyr1278) (D59G10) Rabbit mAb (upper) and ALK (C26G7) Rabbit mAb #3333 (lower).

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).

Phosphorylation of ALK on Tyr1278 was identified at Cell Signaling Technology using PTMScan® technology, a proprietary LC-MS/MS method for phosphorylation site discovery. Phosphorylation of ALK at Tyr1278 was observed in select carcinoma cell lines and in tumors (6).

  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