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  • 研究・開発
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  • Cellular Internalization Kits and Reagents
  • pHSense Reagents
  • pHSense Eu Anti-HA, 10 x 96 wells

pHSense Eu Anti-HA, 10 x 96 wells

Cellular Internalization Kits and Reagents
pHSense Eu Anti-HA, 10 x 96 wells
Cellular Internalization Kits and Reagents
Cellular Internalization Kits and Reagents
Cellular Internalization Kits and Reagents

The pHSense™ Eu Anti-HA carries a pH-sensitive probe and is designed for the detection of HA-tagged receptor and membrane protein internalization.

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Feature Specification
Application 内部化
Sample Volume 50 µL

The pHSense™ Eu Anti-HA carries a pH-sensitive probe and is designed for the detection of HA-tagged receptor and membrane protein internalization.

View product information
Product variants
Unit Size: 96 wells
Part #:
81HAEU1AA
Unit Size: 2 x 96 wells
Part #:
81HAEU1AB
Unit Size: 10 x 96 wells
Part #:
81HAEU1AC
For research use only. Not for use in diagnostic procedures. All products to be used in accordance with applicable laws and regulations including without limitation, consumption and disposal requirements under European REACH regulations (EC 1907/2006).
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Cellular Internalization Kits and Reagents
pHSense Eu Anti-HA, 10 x 96 wells
Cellular Internalization Kits and Reagents
Cellular Internalization Kits and Reagents
pHSense Eu Anti-HA, 10 x 96 wells
Cellular Internalization Kits and Reagents

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Product information

  • Overview
  • How it works
  • Assay validation
  • Specifications

Overview

pHSense Eu Anti-HA is non cell permeant, and labeled with a pH sensitive europium complex. It is specifically designed to monitor the internalization of receptors and membrane proteins tagged with an HA epitope on their extracellular domain. This mouse monoclonal anti-HA antibody (IgG2b) is labeled with pHSense and raised against the twelve-amino-acid peptide sequence CYPYDVPDYASL. It recognizes the influenza virus hemagglutinin epitope (YPYDVPDYA), which has been used extensively as a general epitope tag in expression vectors. At neutral extracellular pH (≥7), its fluorescence remains minimal. Once internalized, the pHSense Eu Anti-HA 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-HA 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-HA assay principle

pHSense Eu Anti-HA is non cell permeant, and labeled with a pH sensitive europium complex. This Monoclonal Anti-HA antibody is designed to monitor receptor and membrane protein internalization when tagged with an HA epitope. The antibody recognizes the HA sequence and remains minimally fluorescent at neutral extracellular pH (≥7). Once internalized, the pHSense Eu Anti-HA encounters increasingly acidic compartments such as early and late endosomes and lysosomes, where the europium fluorescent signal becomes progressively stronger.

Image generic R&D attributes assay principle pHSense Anti-Flag
pHSense generic graph

pHSense Eu Anti-HA assay protocol

The assay begins with culturing cells in a 96-well plate. The pHSense Eu Anti-HA 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 a TRF-compatible plate reader.

simplified pathway pHSense Eu anti HA

Assay validation

pHSense-based detection of GLP1R internalization following exendin-4 stimulation

HEK-293 cells were plated in a 96-well white, culture-treated plate and transfected with a GLP1R-HA-Tagged plasmid.

After cell supernatant removal, 40 µL of the pHSense Eu Anti-HA diluted in cell culture medium (DMEM +10%FBS) were added to the cells, followed by a 1 hour incubation at room temperature.

Exendin-4, a GLP1R agonist, was serially diluted in cell culture medium, and 10 µL of each dilution were added to the wells. After incubation at 37°C, the signal was recorded using a TRF-compatible plate reader. As expected, Exendin-4 induced a dose-dependent increase in signal, with an EC50 value consistent with previously reported data.

assay validation activator Exendin-4 pHSense anti-HA

Specifications

Application
Internalization
Brand
pHSense
Detection Modality
pH sensitive dye
Product Group
Fluorescent Reagent
Sample Volume
50 µL
Shipping Conditions
Shipped in Dry Ice
Target
HA
Target Class
Cell surface proteins, antibodies, ADCs
Technology
TRF
Unit Size
10 x 96 wells

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