Immunofluorescent analysis of Insulin Receptor alpha (green) showing staining in the membrane of HepG2 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 # AHR0221) 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 fluorescent 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|
|Host / Isotype||Mouse / IgG2a, kappa|
|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|
|Functional Assay (FN)||Assay Dependent|
|Inhibition Assays (IA)||Assay Dependent|
|Immunoprecipitation (IP)||2µg per mg lysate|
|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 reacts with an epitope at aa 469-592 (exon 7/8). It primarily reacts with human, but also reacts very weakly with cow, pig and sheep.
This antibody can inhibit insulin binding (~80%). It can also be used in a Tyrosine Kinase assay (Ab-mediated capture on microtitre plates).
Without BSA, this antibody can be used as an insulin-like agonist. Without BSA, it can also be used as both a capture and detection antibody in a sandwich ELISA.
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.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.