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  • PhenoVue Human IgG Antibody Internalization Kit (Fluor 647, pH Sensitive) - 10 x 96-wells

PhenoVue Human IgG Antibody Internalization Kit (Fluor 647, pH Sensitive) - 10 x 96-wells

Images of BT-474 cells showing Trastuzumab internalization compared to an IgG1 isotype control at 1 hour and 24 hours.
Quantification of Trastuzumab internalization in BT-474 cells over time.
Dose–response analysis of Trastuzumab and antibody–drug conjugates (ADCs) in BT-474 cells after 24 hours. View All
PhenoVue Human IgG Antibody Internalization Kit (Fluor 647, pH Sensitive) - 10 x 96-wells
Images of BT-474 cells showing Trastuzumab internalization compared to an IgG1 isotype control at 1 hour and 24 hours.
Quantification of Trastuzumab internalization in BT-474 cells over time.
Dose–response analysis of Trastuzumab and antibody–drug conjugates (ADCs) in BT-474 cells after 24 hours.
Images of BT-474 cells showing Trastuzumab internalization compared to an IgG1 isotype control at 1 hour and 24 hours.
Quantification of Trastuzumab internalization in BT-474 cells over time.
Dose–response analysis of Trastuzumab and antibody–drug conjugates (ADCs) in BT-474 cells after 24 hours.
Representative fluorescence images of BT-474 cells showing Trastuzumab internalization compared to an IgG1 isotype control at 1 hour and 24 hours, acquired with the Opera Phenix™ Plus High-Content Screening System using the PhenoVue™ Human IgG Antibody Internalization Kit (Fluor 647, pH-sensitive). Nuclei are shown in blue, lysosomes in green, and the pH-sensitive Fab–Trastuzumab complex in red.
Images of BT-474 cells showing Trastuzumab internalization compared to an IgG1 isotype control at 1 hour and 24 hours.
Quantification of Trastuzumab internalization in BT-474 cells over time.
Dose–response analysis of Trastuzumab and antibody–drug conjugates (ADCs) in BT-474 cells after 24 hours.
Images of BT-474 cells showing Trastuzumab internalization compared to an IgG1 isotype control at 1 hour and 24 hours.
Quantification of Trastuzumab internalization in BT-474 cells over time.
Dose–response analysis of Trastuzumab and antibody–drug conjugates (ADCs) in BT-474 cells after 24 hours.

The PhenoVue™ human IgG antibody internalization kit offers a ready-to-use, reproducible method for studying antibody internalization. The kit contains PhenoVue Fluor 647 pH-sensitive Fab anti-human IgG, which binds with high affinity to the Fc region of antibodies, whether unconjugated or conjugated (including ADCs). This fluorescent Fab remains weakly fluorescent at neutral extracellular pH (≥7), but its signal increases upon entry into acidic intracellular compartments such as early and late endosomes and lysosomes. This pH-responsive behavior enhances the signal-to-background ratio, supporting clear and reliable visualization of internalization events.

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Feature Specification
Quantity 1
Unit Size 10 x 96-wells

The PhenoVue™ human IgG antibody internalization kit offers a ready-to-use, reproducible method for studying antibody internalization. The kit contains PhenoVue Fluor 647 pH-sensitive Fab anti-human IgG, which binds with high affinity to the Fc region of antibodies, whether unconjugated or conjugated (including ADCs). This fluorescent Fab remains weakly fluorescent at neutral extracellular pH (≥7), but its signal increases upon entry into acidic intracellular compartments such as early and late endosomes and lysosomes. This pH-responsive behavior enhances the signal-to-background ratio, supporting clear and reliable visualization of internalization events.

View product information
Product variants
Assay Points: 2 x 96ウェルマイクロプレート
Part #:
PHAINTDR14
Assay Points: 10 x 96-well microplates
Part #:
PHAINTDR15
For research use only. Not for use in diagnostic procedures.
Request more information
Images of BT-474 cells showing Trastuzumab internalization compared to an IgG1 isotype control at 1 hour and 24 hours.
Quantification of Trastuzumab internalization in BT-474 cells over time.
Dose–response analysis of Trastuzumab and antibody–drug conjugates (ADCs) in BT-474 cells after 24 hours.
Images of BT-474 cells showing Trastuzumab internalization compared to an IgG1 isotype control at 1 hour and 24 hours.
Quantification of Trastuzumab internalization in BT-474 cells over time.
Dose–response analysis of Trastuzumab and antibody–drug conjugates (ADCs) in BT-474 cells after 24 hours.
PhenoVue Human IgG Antibody Internalization Kit (Fluor 647, pH Sensitive) - 10 x 96-wells
Images of BT-474 cells showing Trastuzumab internalization compared to an IgG1 isotype control at 1 hour and 24 hours.
Quantification of Trastuzumab internalization in BT-474 cells over time.
Dose–response analysis of Trastuzumab and antibody–drug conjugates (ADCs) in BT-474 cells after 24 hours.
Images of BT-474 cells showing Trastuzumab internalization compared to an IgG1 isotype control at 1 hour and 24 hours.
Quantification of Trastuzumab internalization in BT-474 cells over time.
Dose–response analysis of Trastuzumab and antibody–drug conjugates (ADCs) in BT-474 cells after 24 hours.
Representative fluorescence images of BT-474 cells showing Trastuzumab internalization compared to an IgG1 isotype control at 1 hour and 24 hours, acquired with the Opera Phenix™ Plus High-Content Screening System using the PhenoVue™ Human IgG Antibody Internalization Kit (Fluor 647, pH-sensitive). Nuclei are shown in blue, lysosomes in green, and the pH-sensitive Fab–Trastuzumab complex in red.
PhenoVue Human IgG Antibody Internalization Kit (Fluor 647, pH Sensitive) - 10 x 96-wells
Images of BT-474 cells showing Trastuzumab internalization compared to an IgG1 isotype control at 1 hour and 24 hours.
Quantification of Trastuzumab internalization in BT-474 cells over time.
Dose–response analysis of Trastuzumab and antibody–drug conjugates (ADCs) in BT-474 cells after 24 hours.
Images of BT-474 cells showing Trastuzumab internalization compared to an IgG1 isotype control at 1 hour and 24 hours.
Quantification of Trastuzumab internalization in BT-474 cells over time.
Dose–response analysis of Trastuzumab and antibody–drug conjugates (ADCs) in BT-474 cells after 24 hours.
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Overview

The study of antibody internalization is important in the research and development of antibody-drug conjugates (ADCs), a rapidly expanding class of biopharmaceuticals that link a monoclonal antibody to a cytotoxic payload via a cleavable linker. Upon internalization and lysosomal processing, the payload is released, triggering cell death through mechanisms such as DNA damage or microtubule disruption. Assessment of antibody internalization is therefore critical for the design, optimization, and evaluation of ADCs and related biologics.

The PhenoVue human IgG antibody internalization kit, which includes PhenoVue 488 lysosomal stain and PhenoVue Hoechst 33342 nuclear stain, facilitates no-wash live-cell, time-lapse monitoring of pH sensitive Fab conjugate-antibody complex trafficking into lysosomes.

The probes included in the kit are compatible with standard cell culture conditions and exert minimal impact on cell viability and proliferation, providing reliable results even during extended live-cell imaging studies.

Specifications

Form
Liquid
Application
High Content Imaging
Microscopy
Assay Points
10 x 96-well microplates
Brand
PhenoVue™
Detection Modality
Fluorescence
Product Group
Kit
Quantity
1
Sample Type
Live cells only
Shipping Conditions
Shipped in Dry Ice
Storage Conditions
-16° C or below, protected from light
Target Species
Human
Type
Kit
Unit Size
10 x 96-wells

Image gallery

Images of BT-474 cells showing Trastuzumab internalization compared to an IgG1 isotype control at 1 hour and 24 hours.
Quantification of Trastuzumab internalization in BT-474 cells over time.
Dose–response analysis of Trastuzumab and antibody–drug conjugates (ADCs) in BT-474 cells after 24 hours.
Images of BT-474 cells showing Trastuzumab internalization compared to an IgG1 isotype control at 1 hour and 24 hours.
Quantification of Trastuzumab internalization in BT-474 cells over time.
Dose–response analysis of Trastuzumab and antibody–drug conjugates (ADCs) in BT-474 cells after 24 hours.
PhenoVue Human IgG Antibody Internalization Kit (Fluor 647, pH Sensitive) - 10 x 96-wells
Images of BT-474 cells showing Trastuzumab internalization compared to an IgG1 isotype control at 1 hour and 24 hours.
Quantification of Trastuzumab internalization in BT-474 cells over time.
Dose–response analysis of Trastuzumab and antibody–drug conjugates (ADCs) in BT-474 cells after 24 hours.
Images of BT-474 cells showing Trastuzumab internalization compared to an IgG1 isotype control at 1 hour and 24 hours.
Quantification of Trastuzumab internalization in BT-474 cells over time.
Dose–response analysis of Trastuzumab and antibody–drug conjugates (ADCs) in BT-474 cells after 24 hours.
PhenoVue Human IgG Antibody Internalization Kit (Fluor 647, pH Sensitive) - 10 x 96-wells
Images of BT-474 cells showing Trastuzumab internalization compared to an IgG1 isotype control at 1 hour and 24 hours.
Quantification of Trastuzumab internalization in BT-474 cells over time.
Dose–response analysis of Trastuzumab and antibody–drug conjugates (ADCs) in BT-474 cells after 24 hours.
Images of BT-474 cells showing Trastuzumab internalization compared to an IgG1 isotype control at 1 hour and 24 hours.
Quantification of Trastuzumab internalization in BT-474 cells over time.
Dose–response analysis of Trastuzumab and antibody–drug conjugates (ADCs) in BT-474 cells after 24 hours.

Spectra viewer

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Application Note
Live-cell imaging of ADC internalization and trafficking

Internalization and trafficking to lysosomes are critical steps for the efficacy of antibody-based treatments, especially antibody...

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Product Info
PhenoVue human IgG antibody internalization kit (Fluor 647, pH sensitive) product information sheet
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