Recombinant Human Insulin R/CD220 (aa 28-944) Protein, CF Summary
Details of Functionality
Measured by its binding ability in a functional ELISA. When 15 ng/mL of biotinylated recombinant human Insulin is added to serially diluted Recombinant Human Insulin R/CD220, the concentration of Recombinant Human Insulin R/CD220 that produces 50% of the optimal binding response is 0.03‑0.15 μg/mL.
Source
Mouse myeloma cell line, NS0-derived human Insulin R/CD220 protein His28-Arg750 ( alpha subunit) & Ser751-Lys944 with a C-terminal 10-His tag ( beta subunit)
>90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Note
<0.10 EU per 1 μg of the protein by the LAL method.
Applications/Dilutions
Dilutions
Binding Activity
Theoretical MW
82.9 kDa ( alpha subunit), 22.9 kDa ( beta subunit). Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
SDS-PAGE
122-135 kDa and 33-43 kDa, reducing conditions
Publications
Read Publications using 1544-IR/CF in the following applications:
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.
Buffer
Lyophilized from a 0.2 μm filtered solution in PBS.
Purity
>90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Reconstitution Instructions
Reconstitute at 100 μg/mL in sterile PBS.
Notes
This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.
Alternate Names for Recombinant Human Insulin R/CD220 (aa 28-944) Protein, CF
CD 220
CD220 antigen
CD220
EC 2.7.10
EC 2.7.10.1
HHF5
INSR
Insulin R
insulin receptor
InsulinR
IR
Background
The Insulin Receptor (gene name INSR, designated CD220) is a type I transmembrane glycoprotein in the Insulin/IGF Receptor family of receptor tyrosine kinases that share structural similarity and overlapping intracellular signaling events (1-3). The 1370 amino acid (aa) human Insulin R preproprotein (A isoform) is processed by proteolysis to remove the signal peptide and produce an extracellular alpha portion (aa 28-750), and an extracellular/transmembrane/cytoplasmic beta subunit (aa 751-1370) (4). The extracellular domain (ECD) contains two homologous globular domains separated by a cysteine-rich domain and followed by three fibronectin type III domains. The intracellular region contains insulin-receptor substrate (IRS) docking sites, the kinase domain, and a phosphotyrosine-containing linker region. The human Insulin R ECD shares 96% aa sequence identity with mouse, rat, equine and canine Insulin R. As a result of alternative splicing, two INSR isoforms that differ by the absence (IR-A) or presence (IR-B) of a 12 aa residue sequence in the carboxyl terminus of the alpha subunit exist (4). IR-A expression is highest in fetal tissues and cancer cells, while IR-B is concentrated in adult differentiated cells (2-5). IR-A and IR-B may homodimerize, or heterodimerize with the IGF-I receptor (1, 3, 4). All receptor combinations bind insulin, IGF-I or IGF-II, but with differing affinities; for example, IR-A has considerably higher affinity for IGF-II as compared to IR-B (2-5). This system allows fine tuning of signaling pathways according to the concentrations of insulin, IGF-I and IGF-II, and expression of receptor subunits on the cell surface (2, 3). Insulin R signaling regulates glucose uptake and metabolism, but also contributes to cell growth, differentiation and apoptosis (2, 3, 5, 6). Mutations in the Insulin R gene have been linked severe insulin resistance (type A and Rabson-Mendenhall syndrome) that may include type II diabetes mellitus and, rarely, leprechaunism (Donohue syndrome) that also includes growth delays and endocrine system abnormalities (1, 7). The R&D Systems human Insulin R consists of the entire ECD of the IR-A isoform.
Nakae, J. et al. (2001) Endoc. Rev. 22:818.
Sciacca, L. et al. (2003) Endocrinology 144:2650.
Belfiore, A. et al. (2009) Endocrine Rev. 30:586.
Lawrence, M.C. et al. (2007) Curr. Opin. Struct. Biol. 17:699.
Sacco, A. et al. (2009) Endocrinology 150:3594.
Kitamura, T. et al. (2004) J. Clin. Invest. 113:209.
Musso, C. et al. (2004) Medicine (Baltimore) 83:209.
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