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7159
PathScan® Phospho-ALK (Tyr1586) Sandwich ELISA Kit
ELISA Kits
ELISA Kit

PathScan® Phospho-ALK (Tyr1586) Sandwich ELISA Kit #7159

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PathScan® Phospho-ALK (Tyr1586) Sandwich ELISA Kit: Image 1

Figure 1: Phosphorylation of ALK and NPM-ALK is detected by PathScan® Phospho-ALK (Tyr1586) Sandwich ELISA Kit #7159 (top, right) in Karpas299 cells lysed in the presence of phosphatase inhibitors (phospho-lysate). In contrast, a low level of phospho-ALK is detected in Karpas299 cells lysed in the absence of phosphatase inhibitors (nonphospho-lysate). Similar levels of total ALK from either nonphospho- or phospho-lysates are detected by PathScan® Total ALK Sandwich ELISA Kit #7322 (top, left). Absorbance at 450 nm is shown in the top figure, while the corresponding Western blots, using Phospho-ALK (Tyr1586) (3B4) Rabbit mAb #3348 (right) or a ALK (C26G7) Rabbit mAb #3333 (left) are shown in the bottom figure. Cell Line Source: Dr Abraham Karpas at the University of Cambridge.

PathScan® Phospho-ALK (Tyr1586) Sandwich ELISA Kit: Image 2

Figure 2: The relationship between protein concentration of phospho- or nonphospho-lysates and the absorbance at 450 nm is shown. Karpas299 cells were harvested at 106 cells/ml, and lysed with or without addition of phosphatase inhibitors to the lysis buffer. Cell Line Source: Dr Abraham Karpas at the University of Cambridge.

Inquiry Info.# 7159

Important Ordering Details

Custom Ordering Details: When ordering five or more kits, please contact us for processing time and pricing at [email protected].

Supporting Data

REACTIVITY

Application Key:

  • W-Western
  • IP-Immunoprecipitation
  • IHC-Immunohistochemistry
  • ChIP-Chromatin Immunoprecipitation
  • IF-Immunofluorescence
  • F-Flow Cytometry
  • E-P-ELISA-Peptide

Species Cross-Reactivity Key:

  • H-Human
  • M-Mouse
  • R-Rat
  • Hm-Hamster
  • Mk-Monkey
  • Vir-Virus
  • Mi-Mink
  • C-Chicken
  • Dm-D. melanogaster
  • X-Xenopus
  • Z-Zebrafish
  • B-Bovine
  • Dg-Dog
  • Pg-Pig
  • Sc-S. cerevisiae
  • Ce-C. elegans
  • Hr-Horse
  • All-All Species Expected

Product Description

CST's PathScan® Phospho-ALK (Tyr1586) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of ALK and the NPM-ALK fusion protein when phosphorylated at Tyr1586. A phospho-ALK (Tyr1586) Rabbit Antibody* has been coated onto the microwells. After incubation with cell lysates, phospho-ALK and phospho-NPM-ALK are captured by the coated antibody. Following extensive washing, an ALK Mouse Antibody* is added to detect the captured phospho-ALK and phospho-NPM-ALK fusion protein. Anti-mouse IgG, HRP-linked Antibody #7076* is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of absorbance for this developed color is proportional to the quantity of ALK and NPM-ALK fusion protein phosphorylated at Tyr1586.

* Antibodies in kit are custom formulations specific to kit.

MW (kDa) 200, 80

Specificity / Sensitivity

CST's PathScan® Phospho-ALK (Tyr1586) Sandwich ELISA Kit #7159 detects endogenous levels of ALK and the NPM-ALK fusion protein when phosphorylated at Tyr1586. High levels of phospho-ALK (Tyr1586) and the phospho-NPM-ALK fusion protein are detected in Karpas299 cells where ALK and NPM-ALK are constitutively phosphorylated (Figure 1). These high levels are abolished in nonphospho-lysates from Karpas299 cells when lysed without the addition of phosphatase inhibitors* to the lysis buffer. Total ALK (phospho and nonphospho) detected by PathScan® Total ALK Sandwich ELISA Kit #7322 and Western analysis, remain unchanged (Figure 1).

* Phosphatase inhibitors includes sodium pyrophosphate, β-glycerophosphate and Na3VO4.

This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

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).

  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.

Pathways & Proteins

Explore pathways + proteins related to this product.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST's products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST's Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

For Research Use Only. Not For Use In Diagnostic Procedures.
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PathScan is a trademark of Cell Signaling Technology, Inc.