Revvity, 64HIFPEG, HTRF Human and Mouse HIF-1α Detection Kit, 500 Assay Points

Overview
HIF-1a is one of the 2 subunits of the transcription factor hypoxia-inducible factor-1 (HIF-1). HIF-1 acts as a master regulator of cellular and systemic response to hypoxia by activating the translation of genes involved in energy metabolism, angiogenesis, and apoptosis, among others. High levels of HIF-1 have been associated with aggressive tumor progression. HIF-1a is also strongly linked to diabetes through its involvement in the glucose uptake of target cells. It has also been implicated in diabetes complications, such as diabetic nephropathy or diabetic wound repair.
How it works
Assay principle
The HIF-1a HTRF assay is a sandwich immunoassay involving two specific anti-human anti-HIF-1a antibodies, respectively labelled with Europium Cryptate (donor) and d2 (acceptor). The intensity of the signal is proportional to the concentration of anti-HIF-1a present in the sample. The detection reagents may be pre-mixed and added in a single dispensing step for direct detection. No washing is needed at any step. After cell lysis, HIF-1a can be detected using the HTRF HIF-1a cellular assay kit reagents and most TR-FRET multimode plate readers.

 

Assay Protocol
Two-plate assay protocol Cells are plated, stimulated, and lysed in the same 96-well culture plate. Lysates are then transferred to the assay plate for the detection of HIF-1a by HTRF reagents. This protocol enables the cells' viability and confluence to be monitored. The antibodies labelled with HTRF fluorophores may be pre-mixed and added in a single dispensing step to further streamline the assay procedure. The assay detection can be run in 96- to 384-well plates by simply resizing each addition volume proportionally. Application-Protein Quantification
Brand-HTRF
Detection Modality-HTRF
Product Group-Kit
Sample Volume-16 µL
Shipping Conditions-Shipped in Dry Ice
Target Class-Biomarkers
Target Species-Human Mouse
Technology-TR-FRET
Therapeutic Area-Metabolism/Diabetes NASH/Fibrosis Oncology & Inflammation
Unit Size-500 assay points