Product Pathways - PathScan ELISA
PathScan® Phospho-ALK (Tyr1604) Chemiluminescent Sandwich ELISA Kit #7020
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|Kit Includes||Volume||Solution Color|
|Phospho-ALK (Tyr1604) Antibody Coated Microwells||96 tests|
|ALK Detection Ab||5.5 ml||Green|
|Anti-mouse IgG, HRP-linked Antibody||5.5 ml||Red|
|Luminol/Enhancer Solution||3 ml||Colorless|
|Stable Peroxide Buffer||3 ml||Colorless|
|Sealing Tape||2 sheets|
|ELISA Wash Buffer (20X)||25 ml||Colorless|
|Sample Diluent||25 ml||Blue|
|Cell Lysis Buffer (10X) #9803||15 ml||Yellowish|
Note: 12 8-well modules –Each module is designed to break apart for 8 tests.
Note: Kit should be stored at 4°C with the exception of Cell Lysis Buffer, which is stored at –20°C (packaged separately).
Species enclosed in parentheses are predicted to react based on 100% sequence homology.
The PathScan® Phospho-ALK (Tyr1604) Chemiluminescent Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of phospho-ALK (Tyr1604), phospho-EML4-ALK or phospho-NPM-ALK fusion proteins with a chemiluminescent readout. Chemiluminescent ELISAs often have a wider dynamic range and higher sensitivity than conventional chromogenic detection. This chemiluminescent ELISA, which is offered in low volume microplates, shows increased signal and sensitivity while using a smaller sample size. A phospho-ALK (Tyr1604) rabbit antibody has been coated onto the microwells. After incubation with cell lysates, only phospho-ALK (Tyr1604) and phospho-ALK fusion proteins are captured by the coated antibody. Following extensive washing, an ALK mouse mAb is added to detect the captured phospho-ALK (Tyr1604) and phospho-ALK fusion proteins. Anti-mouse IgG, HRP-linked antibody is then used to recognize the bound detection antibody. Chemiluminescent reagent is added for signal development. The magnitude of light emission, measured in relative light units (RLU), is proportional to the quantity of phospho-ALK (Tyr1604) or phospho-ALK fusion proteins.Antibodies in kit are custom formulations specific to kit.
Specificity / Sensitivity
PathScan® Phospho-ALK (Tyr1604) Chemiluminescent Sandwich ELISA Kit #7020 detects endogenous levels of phospho-ALK (Tyr1604) protein, phospho-EML4-ALK and phospho-NPM-ALK fusion proteins in human cells. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.
Relationship between protein concentrations of lysates prepared using NCI-H2228 cells lysed with (phospho) and without (nonphospho) the addition of phosphatase inhibitors to the lysis buffer. Graph inset corresponding to the shaded area shows high sensitivity and a linear response at the low protein concentration range.
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).
- 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.
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For Research Use Only. Not For Use In Diagnostic Procedures.