Immunofluorescent analysis of Insulin Receptor alpha (green) showing staining in the cytoplasm and membrane of MCF-7 cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with an Insulin Receptor alpha monoclonal antibody (Product # AHR0231) in 3% BSA-PBS at a dilution of 1:20 and incubated overnight at 4 °C in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a flourescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.
|Tested species reactivity||Bovine, Human, Sheep, Pig, Rabbit|
|Host / Isotype||Mouse / IgG1|
|Immunogen||IM-9 lymphocytes followed by purified insulin receptor.|
|Storage buffer||PBS, pH 7.4, with 0.2% BSA|
|Contains||0.09% sodium azide|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|ELISA (ELISA)||Assay Dependent|
|Flow Cytometry (Flow)||Assay-Dependent|
|Functional Assay (FN)||Assay Dependent|
|Immunohistochemistry (Frozen) (IHC (F))||2-4µg/ml|
|Immunohistochemistry (Paraffin) (IHC (P))||Assay-Dependent|
|Western Blot (WB)||Assay Dependent|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
This antibody is specific for IR and shows no cross-reactivity with insulin-like growth factor (IGF)-receptors. The epitope for this monoclonal antibody is conformational and is located in exon 3.
Staining of formalin-fixed, paraffin tissues requires digestion of tissue sections with pepsin at 1mg/mL in Tris-HCl, pH 2.0, for 15 min at room temperature or 10 min at 37°C. Recommended positive controls include IM-9 lymphocytes, placenta, or breast carcinoma.
Biological actions of insulin and IGF-1 are mediated by their respective cell surface receptor tyrosine kinases that regulate multiple signaling pathways through activation of a series of phosphorylation cascades. The insulin receptor. Insulin/IGF-1 binding to the extracellular domain leads to autophosphorylation of downstream target proteins. These two receptors differ in sequence in regions that confer specificity for the designated ligand as well as in certain intracellular signaling domains, resulting in significant differences in the functional consequences of activation of each receptor. Defects in IR are the cause of various insulin resistance syndromes and IGF-1R defects may cause some forms of growth retardation. Both these signaling cascades are also important for the development of cancer.
IP-MS enrichment of INSR (LFQ intensity): INSR was enriched 904-fold from HCT116 lysate compared to background proteins, using the optimized IP-MS workflow with Pierce MS-Compatible Magnetic IP Kit protein A/G (Part No. 90409) and INSR antibody (Part No. AHR0231). The STRING database (www.string-db.org) was used to identify the protein interactor list. See more information on IP-MS verification of antibody selectivity. IP-MS validation info.
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