Recombinant Human ALK/CD246 His-tag Protein, CF Summary
Product Specifications
Val19-Ser1038, with a C-terminal 6-His tag
Analysis
Product Datasheets
Carrier Free
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.
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.
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Scientific Data
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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
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).
- Chiarle, R. et al. (2008) Nat. Rev. Cancer 8:11.
- Allouche, M. (2007) Cell Cycle 6:1533.
- Iwahara, T. et al. (1997) Oncogene 14:439.
- Morris, S.W. et al. (1997) Oncogene 14:2175.
- Vernersson, E. et al. (2006) Gene Expr. Patterns 6:448.
- Morris, S.W. et al. (1994) Science 263:1281.
- Mourali, J. et al. (2006) Mol. Cell. Biol. 26:6209.
- Moog-Lutz, C. et al. (2005) J. Biol. Chem. 280:26039.
- Motegi, A. et al. (2004) J. Cell Sci. 117:3319.
- Stoica, G.E. et al. (2001) J. Biol. Chem. 276:16772.
- Stoica, G.E. et al. (2002) J. Biol. Chem. 277:35990.
- Mi, R. et al. (2007) Proc. Natl. Acad. Sci. 104:4664.
- Fujimoto, J. et al. (1996) Proc. Natl. Acad. Sci. 93:4181.
- Bischof, D. et al. (1997) Mol. Cell. Biol. 17:2312.
FAQs
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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.
Reconstitution Buffers
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