Revvity, 64NFOPEH, HTRF Human and Mouse Total FOXO1 Detection Kit, 10,000 Assay Points

Overview
The Total FoxO1 cellular assay monitors total FoxO1 and is used as a normalization assay with the phospho-FoxO1 kit. FoxO1 is a member of the Forkhead transcription factor FOXO subfamily, highly expressed in insulin-responsive tissues where it regulates glucose/lipid metabolism and stress resistance. It also functions as a tumor suppressor by inhibiting cell proliferation. FoxO1 deregulation plays a critical role in the development of metabolic disorders such as diabetes and NAFLD. Its dysfunction is also linked to various types of cancer.
How it works
Total-FoxO1 assay principle
The Total-FoxO1 assay quantifies the expression level of FoxO1 in a cell lysate. Contrary to Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis or transfer. The Total-FoxO1 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 FoxO1 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 proteins expression under a no-wash assay format.

 

Total-FoxO1 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 FGFR2 HTRF detection reagents. This protocol enables the cells' viability and confluence to be monitored.

 

Total-FoxO1 1-plate assay protocol
Detection of total FGFR2 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. Application-Cell Signaling
Brand-HTRF
Detection Modality-HTRF
Lysis Buffer Compatibility-Lysis Buffer 1 Lysis Buffer 4 Lysis Buffer 5
Molecular Modification-Total
Product Group-Kit
Sample Volume-16 µL
Shipping Conditions-Shipped in Dry Ice
Target Class-Phosphoproteins
Target Species-Human Mouse
Technology-TR-FRET
Therapeutic Area-Metabolism/Diabetes NASH/Fibrosis Oncology & Inflammation
Unit Size-10,000 assay points