Human Insulin R/CD220 Antibody

Catalog # Availability Size / Price Qty
MAB15442-100
MAB15442-SP
Detection of Insulin R/CD220 in NS0 Mouse Cell Line Transfected with Human Insulin R/CD220 by Flow Cytometry.
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Human Insulin R/CD220 Antibody Summary

Species Reactivity
Human
Specificity
Detects human Insulin R/CD220 in direct ELISAs.
Source
Monoclonal Mouse IgG2B Clone # 243522
Purification
Protein A or G purified from hybridoma culture supernatant
Immunogen
Mouse myeloma cell line NS0-derived recombinant human Insulin R/CD220
His28-Lys956
Accession # P06213
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Flow Cytometry
0.25 µg/5 x 106 cells
See below
Immunocytochemistry
8-25 µg/mL
See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Data Examples

Flow Cytometry Detection of Insulin R/CD220 antibody in NS0 Mouse Cell Line Transfected with Human Insulin R/CD220 antibody by Flow Cytometry. View Larger

Detection of Insulin R/CD220 in NS0 Mouse Cell Line Transfected with Human Insulin R/CD220 by Flow Cytometry. NS0 mouse myeloma cell line transfected with human Insulin R/CD220 (filled histogram), or irrelevant transfectant (open histogram) were stained with Mouse Anti-Human Insulin R/CD220 Monoclonal Antibody (Catalog # MAB15442) or Mouse IgG2B Isotype Control (Catalog # MAB0041, data not shown) followed by APC-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B). View our protocol for Staining Membrane-associated Proteins.

Flow Cytometry Detection of Insulin R/CD220 antibody in Human PBMC lymphocytes antibody by Flow Cytometry. View Larger

Detection of Insulin R/CD220 in Human PBMC lymphocytes by Flow Cytometry. Human PBMC lymphocytes were stained with Mouse Anti-Human Insulin R/CD220 Monoclonal Antibody (Catalog # MAB15442, filled histogram) or Mouse IgG2B Isotype Control (Catalog # MAB0041, open histogram) followed by APC-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B). View our protocol for Staining Membrane-associated Proteins.

Immunocytochemistry Insulin R/CD220 antibody in RT-4 Human Cell Line by Immunocytochemistry (ICC). View Larger

Insulin R/CD220 in RT‑4 Human Cell Line. Insulin R/CD220 was detected in immersion fixed RT-4 human urinary bladder transitional cell papilloma cell line using Mouse Anti-Human Insulin R/CD220 Monoclonal Antibody (Catalog # MAB15442) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cell membrane. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

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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Preparation and Storage

Reconstitution
Reconstitute at 0.5 mg/mL in sterile PBS.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
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.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: Insulin R/CD220

The Insulin Receptor (INS R) and insulin-like growth factor-1 receptor (IGF-1 R) constitute a subfamily of receptor tyrosine kinases (1‑4). The two receptors share structural similarity as well as overlapping intracellular signaling events, and are believed to have evolved through gene duplication from a common ancestral gene. INS R cDNA encodes a type I transmembrane single chain preproprotein with a putative 27 amino acid residues (aa) signal peptide. The large INS R extracellular domain is organized into two successive homologous globular domains, which are separated by a Cysteine-rich domain, followed by three fibronectin type III domains. The intracellular region contains the kinase domain sandwiched between the juxtamembrane domain used for docking insulin-receptor substrates (IRS), and the carboxy-terminal tail that contains two phosphotyrosine-binding sites. After synthesis, the single chain INS R precursor is glycosylated, dimerized and transported to the Golgi apparatus where it is processed at a furin-cleavage site within the middle fibronectin type III domain to generate the mature disulfide-linked  alpha 2 beta 2 tetrameric receptor. The alpha subunit is localized extracellularly and mediates ligand binding while the transmembrane beta subunit contains the cytoplasmic kinase domain and mediates intracellular signaling. As a result of alternative splicing, two INS R isoforms (A and B) that differ by the absence or presence, respectively, of a 12 aa residue sequence in the carboxyl terminus of the alpha subunit exist. Whereas the A isoform is predominantly expressed in fetal tissues and cancer cells, the B isoform is primarily expressed in adult differentiated cells. Both the A and B isoforms bind insulin with high-affinity, but the A isoform has considerably higher affinity for IGF‑I and IGF-II. Ligand binding induces a conformational change of the receptor, resulting in ATP binding, autophosphorylation, and subsequent downstream signaling. INS R signaling is important in metabolic regulation, but may also contribute to cell growth, differentiation and apoptosis. Mutations in the INS R gene have been linked to insulin-resistant diabetes mellitus, noninsulin-dependent diabetes mellitus and leprechaunism, an extremely rare disorder characterized by abnormal resistance to insulin that results in a variety of distinguishing characteristics, including growth delays and abnormalities affecting the endocrine system. INS R is highly conserved between species, rat INS R shares 94% and 97% aa sequence homology with the human and mouse receptor, respectively.

References
  1. Nakae, J. et al. (2001) Endoc. Rev. 22:818.
  2. De Meyts, P. and J. Whittaker (2002) Nature Rev. Drug Disc. 1:769.
  3. Kim, J.J. and D. Accili (2002) Growth Hormone and IGF Res. 12:84.
  4. Sciacca, L. et al. (2003) Endocrinology 144:2650.
Long Name
Insulin Receptor
Entrez Gene IDs
3643 (Human); 16337 (Mouse)
Alternate Names
CD 220; CD220 antigen; CD220; EC 2.7.10; EC 2.7.10.1; HHF5; INSR; Insulin R; insulin receptor; InsulinR; IR

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Isotype Controls

Reconstitution Buffers

Secondary Antibodies

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