Description

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). Moreover, ALK was 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 NSCL patients (6-14).

 

REFERENCES  
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.  Takeuchi, K. et al. (2009) Clin Cancer Res 15, 3143-9.
10.  Palmer, R.H. et al. (2009) Biochem J 420, 345-61.
11.  Horn, L. and Pao, W. (2009) J Clin Oncol 27, 4232-5.
12.  Rodig, S.J. et al. (2009) Clin Cancer Res 15, 5216-23.
13.  Mino-Kenudson, M. et al. (2010) Clin Cancer Res 16, 1561-71.
14.  Kwak, E.L. et al. (2010) N Engl J Med 363, 1693-703.

15.  Martelli, M.P. et al.(2009) Am J Pathol 174, 661-70

  
Products are for research use only. They are not intended for human, animal, or diagnostic applications.

Top: Western Blot detection of ALK fusion protein expression using ALK-specific monoclonal (1B9) antibody.
ALK is found as a fusion protein in NSCLC cell lines and is reported to be expressed by H2228 cells with a MW of 90kDa (11, 15).
Lane A: 293 cells transfected with plasmid encoding EML4 (1-200aa)-ALK (1421-1620aa) fusion protein.
Lane B: 293 cells transfected with mock vector
Lane C: H2228 cells with endogenous  EML4-ALK v3, EML4(1-222aa)-ALK(1058-1620aa), fusion protein.
Lane D: H460 cells (negative control cell lysate).
Middle: Immunocytochemical stainings of H2228 cells and H460 (ALK-negative) cells using ALK-specific monoclonal antibody (clone 1B9). Superview’s ALK monoclonal (1B9) antibody (1:200 dilution). 
Bottom: Immunocytochemical stainings of H2228 cells and H460 (ALK-negative) cells using ALK-specific monoclonal antibody (clone 1B9). DAKO ALK antibody product M7195 (1:200 dilution).  
 

Details

Cat.No.:
CC10035
Antigen:
Recombinant human ALK fusion protein (1421-1620aa) expressed in mammalian cells.
Isotype:
Mouse IgG
Species & predicted
species cross-
reactivity ( ):
Human, Mouse, Rat
Applications &
Suggested starting
dilutions:*
WB                  1:1000-1:2000
IP                    n/d
IHC                  1:200
ICC                  1:200
FACS                n/d
Predicted Molecular
Weight of protein:
220 KDa (ALK), 80 KDa (NPM-ALK), 117 KDa (EML4-ALK v1), 90 KDa (EML4-ALK v3)
Specificity/Sensitivity:
Detects endogenous ALK proteins without cross-reactivity with other family members.
Storage:
Store at -20°C, 4°C for frequent use. Avoid repeated freeze-thaw cycles.
*Optimal working dilutions must be determined by end user.

Products

Product Size CAT.# Price Quantity
Anti-ALK: Mouse ALK Antibody Size: 100 ul CAT.#: CC10035 Price: $325.00