The total Insulin Receptor beta assay enables the detection of cellular Insulin Receptor beta and can be used as a normalization assay for the phospho-IR beta kit.
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  • No-wash
  • Low sample consumption
  • Faster and more convenient than ELISA
The total Insulin Receptor beta assay enables the detection of cellular Insulin Receptor beta and can be used as a normalization assay for the phospho-IR beta kit.
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Overview

Total Insulin Receptor-ß assay enables the detection of Total Insulin Receptor-ß. Stimulated by insulin and insulin-like growth factors, the insulin receptor is involved in several pathways such as those for Insulin and FoxO signaling.

Total-Insulin Receptor ß assay principle

The Total-Insulin Receptor ß assay quantifies the expression level of Insulin Receptor ß in a cell lysate. Contrary to Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis or transfer. The Total-Insulin Receptor ß assay uses two labeled antibodies: one coupled to a donor fluorophore, the other to an acceptor. Both antibodies are highly specific for a distinct epitope on the protein. In presence of Insulin Receptor ß in a cell extract, the addition of these conjugates brings the donor fluorophore into close proximity with the acceptor and thereby generates a FRET signal. Its intensity is directly proportional to the concentration of the protein present in the sample, and provides a means of assessing the protein’s expression under a no-wash assay format.

Total-Insulin Receptor ß 2-plate assay protocol

The 2 plate protocol involves culturing cells in a 96-well plate before lysis then transferring lysates to a 384-well low volume detection plate before adding Total Insulin Receptor ß HTRF detection reagents. This protocol enables the cells' viability and confluence to be monitored.

Total-Insulin Receptor ß 1-plate assay protocol

Detection of total Insulin Receptor ß with HTRF reagents can be performed in a single plate used for culturing, stimulation and lysis. No washing steps are required. This HTS designed protocol enables miniaturization while maintaining robust HTRF quality.
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