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Alpha SureFire Ultra Multiplex phospho/TotalInsulin Receptor β - 50,000 Assay Points

The Alpha SureFire® Ultra Multiplex phospho-Insulin Receptor β (Tyr1150/1151) + Total Insulin Receptor β assay kit is used to measure both the phosphorylation (on Tyr1150/1151) and total levels of endogenous Insulin Receptor β in cellular lysates. The assay is an ideal system for the screening of modulators of receptor activation (e.g. agonists and antagonists) as well as agents acting intracellularly, such as small molecule inhibitors of signal transduction. The assay will measure Insulin-1 Receptor β phosphorylation by either recombinant or endogenous receptors, and can be applied to primary cells.

For research use only. Not for use in diagnostic procedures.

Part Number
Unit Size
List Price
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MPSU-PTINR-K500
500 assay points
3362.00 USD
 
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MPSU-PTINR-K10K
10,000 assay points
20300.00 USD
 
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MPSU-PTINR-K50K
50,000 assay points
63800.00 USD
 
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MPSU-PTINR-K-HV
100 assay points
995.00 USD
 
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Overview

The Alpha SureFire Ultra Multiplex p-Insulin Receptor β + Total Insulin Receptor β assay kit is used to measure both phosphorylated and total levels of endogenous Insulin Receptor β in cellular lysates in a multiplexing Alpha no-wash assay. No specially engineered cell lines are required. The 615 nm (Eu) signal corresponds to the phosphorylated Insulin Receptor β analysis, and the 545 nm (Tb) signal corresponds to the total Insulin Receptor β analysis. This kit has been formulated to provide superior signal:background assay windows and to perform without interference in the presence of extraneous antibodies, making it amenable to the study of therapeutic and blocking antibodies.

  • No-wash steps, no separation steps
  • Multiplexing assay
  • Sensitive detection
  • No interference from exogenous antibodies
  • Results in less than 3 hours

In this assay, the Alpha 615 Acceptor bead is coated with the CaptSure™ antibody, which binds the CaptSure-tagged anti-phospho target antibody. The Alpha 545 Acceptor bead is coated with the CaptSure2™ agent, which binds the CaptSure2 tagged anti-total target protein antibody. The Alpha Donor bead binds the biotinylated anti-total target protein antibody.

Specifications

Assay Target Insulin Rec. β
Assay Target Class Protein
Assay Technology Alpha SureFire Ultra Multiplex
Automation Compatible Yes
Detection Method Alpha
Product Brand Name AlphaLISA SureFire Ultra Multiplex
Shipping Condition Blue Ice
Therapeutic Area Metabolic
Unit Size 50,000 assay points
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Application Note

Simultaneous Detection of Total and Phosphorylated Protein in Cellular Kinase Assay Using Alpha SureFire Ultra Multiplex Technology

Cellular kinase signal transduction pathways are involved in the regulation of many important cellular processes such as cell survival, differentiation, and apoptosis. Kinase signaling networks are typically characterized by multiple kinases arranged in cascades containing nodes with feedback loops and crosstalk between pathways. Numerous assay technologies have been developed for studying kinase signaling pathways, and for screening compound libraries in search of agents to modify receptors or kinase activities. The quality of these assays can be impacted by data variability due to cell seeding inhomogeneity or from compound toxicity. A common method for controlling for variability is to normalize the assay signal to a cellular protein whose level does not change as a function of the treatment.

Alpha SureFire® Ultra Multiplex Phospho and Total assays utilize two types of Alpha Acceptor beads to simultaneously measure two signals from each assay well to easily normalize the assay. Here, we demonstrate the benefit and utility of normalizing assay signal of phosphorylated protein levels to total protein levels in two different cellular models: ERK 1/2 phosphorylation in human melanoma A375 cells and AKT 1/2/3 phosphorylation in mouse myoblast C2C12 cells.

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