Recombinant Human ALK/CD246 His-tag Protein, CF

Catalog # Availability Size / Price Qty
4210-CD-100
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Recombinant Human ALK/CD246 His-tag Protein, CF Summary

Product Specifications

Purity
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Level
<0.10 EU per 1 μg of the protein by the LAL method.
Activity
Measured by its binding ability in a functional ELISA. In a Human ALK/CD246 Antibody (Catalog # AF4210) coated plate, when Recombinant Human ALK/CD246 is present at 0.5 μg/mL, Recombinant Human Pleiotrophin/PTN (Catalog # 252-PL) binds with an ED50 of 20-100 ng/mL.
Source
Mouse myeloma cell line, NS0-derived human ALK/CD246 protein
Val19-Ser1038, with a C-terminal 6-His tag
Accession #
N-terminal Sequence
Analysis
Val19
Predicted Molecular Mass
111 kDa
SDS-PAGE
130-150 kDa, under reducing conditions

Product Datasheets

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

4210-CD

Formulation Lyophilized from a 0.2 μm filtered solution in PBS.
Reconstitution Reconstitute at 500 μg/mL in PBS.
Shipping The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.

Data Images

Binding Activity View Larger

In a Human ALK/CD246 Antibody (AF4210) coated plate, when Recombinant Human ALK/CD246 (Catalog # 4210-CD) is present at 0.5 μg/mL, Recombinant Human Pleiotrophin/PTN (252-PL) binds with an ED50 of 20-100 ng/mL.

SDS-PAGE View Larger

2 μg/lane of Recombinant Human ALK.CD246 His-tag (Catalog # 4210-CD) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 130-150 kDa.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: ALK/CD246

Anaplastic Lymphoma Kinase (ALK), also known as CD246, is a transmembrane receptor tyrosine kinase in the Insulin Receptor family. The ALK gene is a target of multiple chromosomal translocations in cancer that encode hybrid proteins which promote cellular transformation (1, 2). Human ALK consists of a 1020 amino acid (aa) extracellular domain (ECD) with two MAM domains that flank an LDLR class A domain, a 21 aa transmembrane segment, and a 561 aa cytoplasmic domain that contains the tyrosine kinase domain (3, 4). Within the ECD, human ALK shares 89% aa sequence identity with mouse and rat ALK. It is primarily expressed in the developing nervous system but is also found in various non‑neural tissues (3 ‑ 6). Mature ALK is expressed on the cell surface as a 200 ‑ 220 kDa N‑glycosylated protein (3, 4, 7). Proteolytic cleavage of ALK liberates an 80 kDa soluble fragment from the ECD with a 140 kDa fragment remaining cell‑associated (8, 9). ALK is classified as a dependence receptor, a protein that promotes apoptosis in the absence of ligand but is anti‑apoptotic upon stimulation (2, 7). Its cytoplasmic domain is cleaved following Asp1160 by Caspases during apoptosis (7). ALK stimulation by antibody ligation induces activation of its kinase domain and receptor phosphorylation, enabling the association of ALK with signal transduction proteins (4, 9). ALK binds the cytokines Pleiotrophin and Midkine, although their effects on cellular responses are not consistent between different systems (8, 10 ‑ 12). ALK promotes neurite formation in neuroblastoma cells and mediates the neuroprotective effects of Pleiotrophin in motor neurons (9, 12). A t(2;5) chromosomal translocation with Nucleophosmin (NPM) in human anaplastic large cell lymphoma results in a hybrid protein consisting of NPM fused to the kinase domain of ALK (6, 13). NPM‑ALK as well as other ALK fusion proteins are aberrantly localized to the cytoplasm and nucleus, have constitutively active kinase domains, and promote cellular transformation (1, 13, 14).

References
  1. Chiarle, R. et al. (2008) Nat. Rev. Cancer 8:11.
  2. Allouche, M. (2007) Cell Cycle 6:1533.
  3. Iwahara, T. et al. (1997) Oncogene 14:439.
  4. Morris, S.W. et al. (1997) Oncogene 14:2175.
  5. Vernersson, E. et al. (2006) Gene Expr. Patterns 6:448.
  6. Morris, S.W. et al. (1994) Science 263:1281.
  7. Mourali, J. et al. (2006) Mol. Cell. Biol. 26:6209.
  8. Moog-Lutz, C. et al. (2005) J. Biol. Chem. 280:26039.
  9. Motegi, A. et al. (2004) J. Cell Sci. 117:3319.
  10. Stoica, G.E. et al. (2001) J. Biol. Chem. 276:16772.
  11. Stoica, G.E. et al. (2002) J. Biol. Chem. 277:35990.
  12. Mi, R. et al. (2007) Proc. Natl. Acad. Sci. 104:4664.
  13. Fujimoto, J. et al. (1996) Proc. Natl. Acad. Sci. 93:4181.
  14. Bischof, D. et al. (1997) Mol. Cell. Biol. 17:2312.
Long Name
Anaplastic Lymphoma Kinase
Entrez Gene IDs
238 (Human); 11682 (Mouse); 266802 (Rat)
Alternate Names
ALK tyrosine kinase receptor; ALK; anaplastic lymphoma kinase (Ki-1); Anaplastic lymphoma kinase; anaplastic lymphoma receptor tyrosine kinase; CD246 antigen; CD246; EC 2.7.10.1; Ki-1; NBLST3; Tcrz

FAQs

  1. Does your Recombinant Human ALK/CD246 His-tag Protein (Catalog # 4210-CD) exhibit kinase activity?

    • Our Recombinant Human ALK/CD246 His-tag Protein (Catalog # 4210-CD) is expressed as the extracellular domain (Val19-Ser1038) of the protein. As the kinase domain is in the cytoplasmic region (Arg1060-Pro1620), we do not expect this protein to exhibit kinase activity.

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