Revvity, 81FL1EU1AC, pHSense Eu Anti-FLAG, 10 x 96 wells

Overview
pHSense™ Eu Anti-FLAG is non cell permeant, and is labeled with a pH sensitive europium complex. This Monoclonal Anti-FLAG antibody is designed to monitor receptor and membrane protein internalization when tagged with the FLAG® sequence (DYKDDDDK) on the extracellular part. The antibody recognizes the FLAG® sequence fused at the N-terminus, intra or C-terminus and remains minimally fluorescent at neutral extracellular pH (≥7). Once internalized, the pHSense™ Eu Anti-FLAG encounters increasingly acidic compartments such as early and late endosomes and lysosomes, where the europium signal becomes progressively stronger. This new pH sensitive europium anti-FLAG 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-Anti-FLAG assay principle
pHSense Eu Anti-FLAG is non cell permeant, and is labeled with a pH sensitive europium complex. This Monoclonal Anti-FLAG antibody is designed to monitor receptor and membrane protein internalization when tagged with the FLAG® sequence. The antibody recognizes the FLAG sequence and remains minimally fluorescent at neutral extracellular pH (≥7). Once internalized, the pHSense Eu Anti-FLAG encounters increasingly acidic compartments such as early and late endosomes and lysosomes, where the europium fluorescent signal becomes progressively stronger.

pHSense Eu Anti-FLAG Assay Protocol
The assay begins by culturing cells in a 96-well plate. The pHSense Eu Anti-FLAG 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-FLAG
Target Class-Cell surface proteins, antibodies, ADCs
Technology-TRF
Unit Size-10 x 96 wells