ALK (D5F3®) XP® Rabbit mAb #3633
- WB
- IP
- IHC
- F
Supporting Data
| REACTIVITY | H |
| SENSITIVITY | Endogenous |
| MW (kDa) | 220 (ALK), 80 (NPM-ALK), 117 (EML4-ALK v1), 86 (EML4-ALK v3) |
| Source/Isotype | Rabbit IgG |
Application Key:
- WB-Western Blotting
- IP-Immunoprecipitation
- IHC-Immunohistochemistry
- F-Flow Cytometry
Species Cross-Reactivity Key:
- H-Human
Product Information
Product Usage Information
| Application | Dilution |
|---|---|
| Western Blotting | 1:2000 |
| Simple Western™ | 1:10 - 1:50 |
| Immunoprecipitation | 1:100 |
| Immunohistochemistry (Paraffin) | 1:125 - 1:500 |
| Flow Cytometry (Fixed/Permeabilized) | 1:400 - 1:800 |
Storage
Protocol
Specificity / Sensitivity
ALK (D5F3®) XP® Rabbit mAb detects endogenous levels of total ALK protein as well as ALK fusion proteins, such as EML4-ALK variants and NPM-ALK, even at low levels. This antibody does not cross-react with other family members.
Species Reactivity:
Human
Source / Purification
Monoclonal antibody is produced by immunizing animals with recombinant protein corresponding to residues in the carboxy terminus of human ALK.
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).
Investigators have identified ALK translocations with other fusion partners, such as TRK-fused gene (TFG) and KIF5B, which have also been associated with NSCLC (6,7). In particular, the EML4-ALK fusion protein has been found in 3-7% of NSCLC samples (6-14).
- Stoica, G.E. et al. (2001) J Biol Chem 276, 16772-9.
- Iwahara, T. et al. (1997) Oncogene 14, 439-49.
- Morris, S.W. et al. (1997) Oncogene 14, 2175-88.
- Morris, S.W. et al. (1994) Science 263, 1281-4.
- Bai, R.Y. et al. (1998) Mol Cell Biol 18, 6951-61.
- Rikova, K. et al. (2007) Cell 131, 1190-203.
- Takeuchi, K. et al. (2008) Clin Cancer Res 14, 6618-24.
- Soda, M. et al. (2007) Nature 448, 561-6.
- Takeuchi, K. et al. (2009) Clin Cancer Res 15, 3143-9.
- Palmer, R.H. et al. (2009) Biochem J 420, 345-61.
- Horn, L. and Pao, W. (2009) J Clin Oncol 27, 4232-5.
- Rodig, S.J. et al. (2009) Clin Cancer Res 15, 5216-23.
- Mino-Kenudson, M. et al. (2010) Clin Cancer Res 16, 1561-71.
- Kwak, E.L. et al. (2010) N Engl J Med 363, 1693-703.
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