Phospho-ALK (Tyr1096) Antibody

4143:

Immunoprecipitation , Western Blotting

Phospho-ALK (Tyr1096) Antibody detects ALK only when phosphorylated at Tyr1096, which is equivalent to Tyr156 of NPM-ALK. This antibody may also cross-react with other overexpressed tyrosine phosphorylated proteins.

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr1096 of human ALK. Antibodies are purified by protein A and peptide affinity chromatography

Western Blotting

Western blot analysis of extracts from KARPAS-299 cells, untreated or treated with calf intestinal phosphatase (CIP), using Phospho-ALK (Tyr1096) Antibody (upper) and ALK (C26G7) Rabbit mAb #3333 (lower).

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 Tyr1096 was identified at Cell Signaling Technology (CST) using PhosphoScan^® , CST's LC-MS/MS platform for phosphorylation site discovery. Phosphorylation of fusion protein NPM-ALK at the Tyr1096 site was also reported by several other labs in select carcinoma cell lines and in tumors and shown to be important for NPM-ALK function (8,9).

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.
9. Turner, S.D. et al. (2007) Cell Signal 19, 740-7.
10. Chikamori, M. et al. (2007) Oncogene 26, 2950-4.

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• 8112 SignalStain^® Antibody Diluent
• 8114 SignalStain^® Boost IHC Detection Reagent (HRP, Rabbit)
• 3333 ALK (C26G7) Rabbit mAb
• 3341 Phospho-ALK (Tyr1604) Antibody
• 3343 Phospho-ALK (Tyr1586) Antibody
• 3710 Phospho-ALK (Tyr1278) Antibody
• 3714 Phospho-ALK (Tyr1282/1283) Antibody
• 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.