Phospho-Insulin/IGF Receptor Antibody Sampler Kit #8338
Product Information
Kit Usage Information
Protocols
Product Description
The Phospho-Insulin/IGF Receptor Antibody Sampler Kit provides an economical means of evaluating total Insulin Receptor and IGF-I Receptor β protein levels as well as Insulin and IGF-I Receptor β phosphorylated at specific sites. The kit includes enough antibody to perform two western blot experiments with each primary antibody.
Specificity / Sensitivity
Each antibody in the Phospho-Insulin/IGF Receptor Antibody Sampler Kit recognizes endogenous levels of the specific target protein, with activation state-specific antibodies recognizing target proteins only when phosphorylated at the indicated residues. IGF-I Receptor ß (D23H3) XP® Rabbit mAb detects endogenous levels of total IGF-I receptor ß protein. This antibody does not cross-react with insulin receptor.Phospho-IGF-I Receptor beta (Tyr1135) (DA7A8) Rabbit mAb detects endogenous levels of IGF-I receptor only when phosphorylated at Tyr1135. This antibody cross-reacts with Tyr1150 of insulin receptor and may also cross-react with other overexpressed related tyrosine-phosphorylated tyrosine kinases. Phospho-IGF-I Receptor ß (Tyr1131)/Insulin Receptor ß (Tyr1146) Antibody detects endogenous levels of Tyr1131-phosphorylated IGF-I receptor and Tyr1146-phosphorylated insulin receptor. This antibody cross-reacts with activated PDGF, FGF and EGF receptors; ErbB2; and c-Met. Phospho-IGF-I Receptor ß (Tyr1135/1136)/Insulin Receptor ß (Tyr1150/1151) (19H7) Rabbit mAb detects endogenous levels of IGF-I receptor and insulin receptor only when phosphorylated at tyrosine 1135/1136 or tyrosine 1150/1151, and does not cross-react with other related tyrosine-phosphorylated tyrosine kinases. Phospho-IGF-I Receptor ß (Tyr980) (C14A11) Rabbit mAb detects endogenous levels of IGF-I receptor ß protein only when phosphorylated at Tyr980, and may cross-react with activated insulin receptors and FLT3. Insulin Receptor ß (4B8) Rabbit mAb detects endogenous levels of total insulin receptor ß and does not cross-react with IGF-I receptor ß.
Source / Purification
Monoclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to the carboxy-terminal residues of human IGF-IR β or residues surrounding Tyr960 of human insulin receptor β. Activation state-specific monoclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr1135 of human IGF-I Receptor β, Tyr1135/1136 of human IGF-I Receptor β, or Tyr980 of human IGF-I Receptor β. Activation state-specific polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues of human IGF-I Receptor β. Polyclonal antibodies are purified by protein A and peptide affinity chromatography.
Background
Type I insulin-like growth factor receptor (IGF-IR) is a transmembrane receptor tyrosine kinase that is widely expressed in many cell lines and cell types within fetal and postnatal tissues (1-3). Receptor autophosphorylation follows binding of the IGF-I and IGF-II ligands. Three tyrosine residues within the kinase domain (Tyr1131, Tyr1135, and Tyr1136) are the earliest major autophosphorylation sites (4). Phosphorylation of these three tyrosine residues is necessary for kinase activation (5,6). Insulin receptors (IRs) share significant structural and functional similarity with IGF-I receptors, including the presence of an equivalent tyrosine cluster (Tyr1146/1150/1151) within the kinase domain activation loop. Tyrosine autophosphorylation of IRs is one of the earliest cellular responses to insulin stimulation (7). Autophosphorylation begins with phosphorylation at Tyr1146 and either Tyr1150 or Tyr1151, while full kinase activation requires triple tyrosine phosphorylation (8).
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