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PathScan® Total ALK Chemiluminescent Sandwich ELISA Kit #7084

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ELISA Image 1: PathScan® Total ALK Chemiluminescent Sandwich ELISA Kit
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.
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To Purchase # 7084**

Important Ordering Details

Custom Ordering Details:

If kit quantities from the same lot are needed in unlisted sizes, contact us for processing time and pricing.

Looking for this ELISA kit in a 384-well format? Inquire for availability, processing time, and pricing.

Supporting Data

REACTIVITY H
Application Key:
  • ELISA-ELISA 
Species Cross-Reactivity Key:
  • H-Human 
  • Product Includes
  • Related Products
Product IncludesVolumeSolution Color
ALK Rabbit mAb Coated Microwells #1990096 tests
ALK Mouse Detection mAb #139931 eaGreen (Lyophilized)
Anti-mouse IgG, HRP-linked Antibody (ELISA Formulated) #133041 eaRed (Lyophilized)
Detection Antibody Diluent #133395.5 mlGreen
HRP Diluent #135155.5 mlRed
Luminol/Enhancer Solution #848503 ml
Stable Peroxide Buffer #425523 ml
Sealing Tape #545032 ea
ELISA Wash Buffer (20X) #980125 ml
ELISA Sample Diluent #1108325 mlBlue
Cell Lysis Buffer (10X) #980315 ml

Kit contents scale proportionally with size, except sealing tape.
Example: The V1 kit contains 5X the listed quantities above, but will exclude the sealing tape.

The microwell plate is supplied as 12 8-well modules - Each module is designed to break apart for 8 tests.

Product Information

Product Description

The PathScan® Total ALK Chemiluminescent Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of total ALK protein and EML4-ALK or NPM-ALK fusion proteins with a chemiluminescent readout. Chemiluminescence 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. An ALK rabbit antibody has been coated onto the microwells. After incubation with cell lysates, ALK and ALK fusion proteins are captured by the coated antibody. Following extensive washing, an ALK mouse antibody is added to detect the captured ALK and 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 magnitude of light emission, measured in relative light units (RLU) is proportional to the quantity of total ALK and ALK fusion proteins.

*Antibodies in this kit are custom formulations specific to kit.

Protocol

Specificity / Sensitivity

PathScan® Total ALK Chemiluminescent Sandwich ELISA Kit #7084 detects endogenous levels of total ALK, EML4-ALK and 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.

Species Reactivity:

Human

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).

Pathways

Explore pathways related to this product.


Database Links

UniProt ID: Q9UM73


Entrez-Gene Id: 238


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For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
PathScan is a registered trademark of Cell Signaling Technology, Inc.
U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.
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