Phospho-Insulin Receptor (Tyr1150/1151) & Total Insulin Receptor Cellular Assay Kits

Phospho- and total Insulin Receptor assays for IR detection

Cisbio's cell-based homogeneous kit for phospho-Insulin receptor (Tyr1150/1151) enables the quantitative detection of Insulin Receptor modulation, phosphorylated at Tyrosines 1150 & 1151. The HTRF® total Insulin Receptor assay kit is designed for the quantitative detection of total Insulin Receptor, phosphorylated and unphosphorylated, to normalize the phosphorylation status of Insulin Receptor with respect to its steady-state level in cells. The buffers of both HTRF® phospho- and total IR assays are compatible, enabling an analysis of the phosphorylated and the total protein populations from one lysate sample. Stimulated by insulin and insulin-like growth factors, the insulin receptor is involved in several pathways such as those for Insulin and FoxO signaling. This tyrosine kinase receptor acts as one of the major receptors in glycemia regulation, and is a pertinent protein in the diabetes and metabolism research areas. The simple add-and-read protocol eliminates all wash steps for a faster analysis and easy miniaturization, while maintaining a high-quality specific output and offering enhanced convenience over ELISA or WB.

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Assay principle

HTRF® - the homogeneous cell-based sandwich immunoassay

Cisbio Bioassay’s phospho- and total IR assays are based on a TR-FRET sandwich immunoassay format comprising two specific anti-IR antibodies, one labeled with Eu3+-cryptate (donor) and the other with d2 (acceptor). The antibodies specifically bind with IR, and the proximity of donor and acceptor will then lead to a fluorescent TR-FRET signal. The protocol is optimized for a 384-well plate format, but can easily be further miniaturized or upscaled. Only low sample volumes are needed. The detection reagents may be pre-mixed and added in a single dispensing step for direct detection. No washing is needed at any step.
The assays can be run with frozen cell lysates or fresh cells in culture. After cell lysis, endogenous phospho- or total-IR can be quantitatively detected using the HTRF phospho-IR (Tyr1150/1151) and total IR cellular assay kit reagents and most TR-FRET multimode plate readers.

A simpler, more flexible assay protocol - adapted to your applications

Two-plate assay protocol

For added flexibility, the assay can be run under a two-plate assay protocol, where cells are plated and treated in a 96-well culture plate. For detection, lysates are subsequently transferred to a 384sv assay plate where the HTRF® reagents are added. This also enables monitoring of the cells' viability and confluence in an appropriate cell culture plate.

What to expect at the bench

This video covers the principles of phospho-HTRF assays and gives guidelines for performing them, using our phospho-ERK assay as an example.

Part#, inserts & MSDS

Ordering Info

DescriptionCat. noProduct insertMSDS
IR beta phospho-Y1150 /1151 kit - 500 tests63ADK016PEG
IR beta phospho-Y1150 /1151 kit - 10,000 tests63ADK016PEH
IR beta phospho-Y1150 /1151 kit - 50,000 tests63ADK016PEY
IR beta total kit - 500 tests63ADK019PEG
IR beta total kit - 10,000 tests63ADK019PEH
IR beta total kit - 50,000 tests63ADK019PEY

Companion products

DescriptionCat. noProduct insertMSDS
IR beta phospho-Y1150/ 1151 kit - control lysate63ADK016TDA
IR beta total kit - control lysate63ADK019TDA
Phospho-total protein blocking reagent - 2 ml64KB1AAC
Phospho-total protein blocking reagent - 6 ml64KB1AAD
Phospho-total protein lysis buffer #1 - 130 mL64KL1FDF