Phospho-ALK (Tyr1282/1283) (D3

9687:

Immunoprecipitation , Western Blotting

Phospho-ALK (Tyr1282/1283) (D39B2) Rabbit mAb detects ALK only when phosphorylated at Tyr1282/1283 (equivalent to Tyr342/343 of NPM-ALK). The antibody may cross-react with other overexpressed phospho-tyrosine proteins such as EGFR.

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr1282/1283 of human ALK protein.

Western Blotting

Western blot analysis of extracts from SUP-M2 cells, untreated or treated with calf intestinal phosphatase (CIP), using Phospho-ALK (Tyr1282/1283) (D39B2) Rabbit mAb (upper) or ALK (C26G7) Rabbit mAb #3333 (lower). The lower molecular weight cluster is partial degradation of the ALK fusion.

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 at Tyr1282/1283 was identified at Cell Signaling Technology (CST) using PhosphoScan^® , CST's LC-MS/MS platform for phosphorylation site discovery. Phosphorylation of ALK at Tyr1282/1283 was observed in select carcinoma cell lines and 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.

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

• 7071 Phototope^® -HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
• 7074 Anti-rabbit IgG, HRP-linked Antibody
• 7720 Prestained Protein Marker, Broad Range (Premixed Format)
• 7727 Biotinylated Protein Ladder Detection Pack
• 7003 20X LumiGLO^® Reagent and 20X Peroxide

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