Anti-Insulin Receptor alpha antibody [83-7] (ab36550)
- Datasheet
- Specific References (5)
- Protocols
Overview
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Product nameAnti-Insulin Receptor alpha antibody [83-7]
See all Insulin Receptor alpha primary antibodies -
DescriptionMouse monoclonal [83-7] to Insulin Receptor alpha
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Host speciesMouse
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SpecificityDoes not cross react with the Human Type 1 IGF Receptor.
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Tested applicationsSuitable for: Flow Cyt, IP, ELISA, IHC-Pmore details
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Species reactivityReacts with: Human
Does not react with: Rat -
Immunogen
Transfected 3T3 cells overexpressing the human Insulin Receptor alpha
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Epitopeab36550 recognizes an epitope within amino acids 140-301 (the cysteine rich region)of the extracellular domain of the human Insulin Receptor alpha.
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General notes
The antibody enhances the binding of 125I insulin binding to the insulin receptor of HIR3.5/3T3 cells and stimulates insulin mediated 3H thymidine incorporation in these cells.
Properties
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FormLiquid
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Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
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Storage bufferPreservative: None
Constituents: PBS -
Concentration information loading... -
PurityProtein G purified
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Primary antibody notesThe antibody enhances the binding of 125I insulin binding to the insulin receptor of HIR3.5/3T3 cells and stimulates insulin mediated 3H thymidine incorporation in these cells.
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ClonalityMonoclonal
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Clone number83-7
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IsotypeIgG1
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Research areas
Associated products
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Compatible Secondaries
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Conjugation kits
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Immunohistochemistry kits
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Isotype control
Applications
Our Abpromise guarantee covers the use of ab36550 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
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| Flow Cyt | Use 1µg for 106 cells. (paraformaldehyde or methanol fixed cells)
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
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| IP | 1/5000. | |
| ELISA | 1/5000. | |
| IHC-P | Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
Target
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RelevanceThe human insulin receptor is a heterotetrameric membrane glycoprotein consisting of disulfide linked subunits in a beta-alpha-alpha-beta configuration. The beta subunit (95 kDa) possesses a single transmembrane domain, whereas the alpha subunit (135 kDa) is completely extracellular. The insulin receptor exhibits receptor tyrosine kinase (RTK) activity. RTKs are single pass transmembrane receptors that possess intrinsic cytoplasmic enzymatic activity, catalyzing the transfer of the gamma phosphate of ATP to tyrosine residues in protein substrates. RTKs are essential components of signal transduction pathways that affect cell proliferation, differentiation, migration and metabolism. Included in this large protein family are the insulin receptor and the receptors for growth factors such as epidermal growth factor, fibroblast growth factor and vascular endothelial growth factor. Receptor activation occurs through ligand binding, which facilitates receptor dimerization and autophosphorylation of specific tyrosine residues in the cytoplasmic portion. The interaction of insulin with the alpha subunit of the insulin receptor activates the protein tyrosine kinase of the beta subunit, which then undergoes an autophosphorylation that increases its tyrosine kinase activity. Three adapter proteins, IRS1, IRS2 and Shc, become phosphorylated on tyrosine residues following insulin receptor activation. These three phosphorylated proteins then interact with SH2 domain containing signaling proteins.
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Cellular localizationMembrane; single pass type I membrane protein.
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Database links
- Entrez Gene: 3643 Human
- Omim: 147670 Human
- SwissProt: P06213 Human
- Unigene: 465744 Human
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Alternative names
- CD220 antibody
- HHF5 antibody
- HIR A antibody
see all
Images
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Insulin Receptor alpha antibody [83-7] (ab36550)](https://www.abcam.com/ps/products/36/ab36550/Images/Insulin-Receptor-alpha-Primary-antibodies-ab36550-1.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Insulin Receptor alpha antibody [83-7] (ab36550)ab36550 (1µg/ml) staining insulin receptor alpha in human pancreas using an automated system (DAKO Autostainer Plus). Using this protocol there is strong staining of cytoplasm and basal cell membrane of proximal convoluted tubule cells.
Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer citrate pH 6.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required. -
![Flow Cytometry - Anti-Insulin Receptor alpha antibody [83-7] (ab36550)](https://www.abcam.com/ps/products/36/ab36550/Images/Insulin-Receptor-alpha-Primary-antibodies-ab36550-2.jpg)
Flow Cytometry - Anti-Insulin Receptor alpha antibody [83-7] (ab36550)Overlay histogram showing Jurkat cells stained with ab36550 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab36550, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Jurkat cells fixed with methanol (5 min) used under the same conditions.
Please note that Abcam do not have data for use of this antibody on non-fixed cells. We welcome any customer feedback.
References
This product has been referenced in:
- Subramanian K et al. Insulin receptor-insulin interaction kinetics using multiplex surface plasmon resonance. J Mol Recognit 26:643-52 (2013). Read more (PubMed: 24277609) »
- Lu JZ et al. Expression in CHO cells and pharmacokinetics and brain uptake in the Rhesus monkey of an IgG-iduronate-2-sulfatase fusion protein. Biotechnol Bioeng 108:1954-64 (2011). Read more (PubMed: 21351076) »
Customer reviews and Q&As
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"