Revvity, 81SNEU1AB, pHSense Eu SNAP Labeling Reagent, 2 x 96 wells

Overview
The pHSense™ Eu SNAP Labeling Reagent is non cell permeant, and is labeled with a pH sensitive europium complex that specifically and covalently interacts with SNAP-tag® receptors and membrane proteins to monitor their internalization. Cells expressing receptors and membrane proteins tagged with a SNAP-tag on the extracellular part can be labeled with pHSense™ Eu SNAP Labeling Reagent and remain minimally fluorescent at neutral extracellular pH (≥7). Once internalized, the pHSense™ Eu SNAP Labeling Reagent encounters increasingly acidic compartments such as early and late endosomes and lysosomes, where the europium signal becomes progressively stronger. This new pH sensitive europium SNAP-labeling reagent is compatible with a time-resolved fluorescence (TRF) detection, effectively eliminating most fluorescence background and significantly enhancing the signal-to-background ratio. Its unique photophysical properties enable simple and robust no-wash detection of receptor-mediated endocytosis in plate-based assays with live-cells.
How it works
pHSense Eu SNAP labeling reagent assay principle
pHSense Eu SNAP Labeling Reagent is non cell permeant, and is labeled with a pH sensitive europium complex that specifically and covalently interacts with SNAP-tag® receptors and membrane proteins designed to monitor their internalization. Cells expressing receptors and membrane proteins tagged with a SNAP-tag on the extracellular part can be labeled with pHSense Eu SNAP Labeling Reagent, and remain minimally fluorescent at neutral extracellular pH (≥7). Once internalized, the pHSense Eu SNAP Labeling Reagent encounters increasingly acidic compartments such as early and late endosomes and lysosomes, where the europium fluorescent signal becomes progressively stronger.

pHSense Eu SNAP labeling reagent assay protocol
The assay begins by culturing cells in a 96-well plate. The pHSense SNAP labeling reagent is then added to the cells and incubated for 1 hour at room temperature. Following this incubation, cells are stimulated with a pharmacological compound. Fluorescence is then measured either kinetically or at endpoint using an HTRF-compatible plate reader. Application-Internalization
Brand-pHSense
Detection Modality-pH sensitive dye
Product Group-Fluorescent Reagent
Sample Volume-50 µL
Shipping Conditions-Shipped in Dry Ice
Target-SNAP-tag fusion protein
Target Class-Cell surface proteins, antibodies, ADCs
Technology-TRF
Unit Size-2 x 96 wells