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Your Position: ホーム > ADC Internalization Detection Antibody – Labeled with pH-Sensitive Fluorescent Dye

ADC Internalization Detection Antibody – Labeled with pH-Sensitive Fluorescent Dye

ADC Internalization Detection Reagent

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Product List
Product Features
Principles & Applications
Validation Data
Resources
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Background
Antibody-drug conjugates (ADCs) are targeted cancer treatments consisting of a monoclonal antibody linked to a cytotoxic drug. The antibody targets specific cancer cell antigens, allowing the drug to be delivered directly to tumors while reducing off-target effects.[1] A key step in the mechanism of action (MOA) of ADCs is receptor-mediated endocytosis. After binding to the target antigen, the ADC is internalized, forming early endosomes. These endosomes then mature into late endosomes and lysosomes, where the linker is cleaved to release the cytotoxic drug. The drug then disrupts DNA replication or microtubule function, triggering tumor cell apoptosis or death.[2]
Therefore, screening antibodies with high-efficiency internalization capabilities is critical during early-stage development.
Classic mechanism of action of ADC drugs

Classic mechanism of action of ADC drugs

Product List
To support ADC internalization research, we have developed a specialized Antibody Internalization Detection Reagent (Cat. No. IGG-PZF2001), featuring a pH-sensitive red fluorescent dye that labels the human IgG Fc region. The reagent forms a stable fluorescent complex that enables detection of antibody endocytosis within cells. It delivers enhanced fluorescence in acidic compartments with minimal background, supporting flow cytometry, cell imaging, and other cellular analysis techniques.
Cat. No.Product DescriptionTests
IGG-PZF2001Antibody Internalization Detection reagent100tests/500tests

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Product Features
High Signal-to-Noise Ratio: Strong fluorescence with minimal background.
Quick Labeling: Complete in just 10 minutes.
pH-Sensitive: Bright signal in acidic intracellular compartments.
Fab Region Preserved: Not affect antibody binding.
Principles & Applications
Principles
Schematic of antibody internalization detection reagent principle

Schematic of antibody internalization detection reagent principle

Applications
Discovery stage: To identify antibodies that not only bind specifically to tumor-associated antigens but also internalize efficiently.
Lead optimization: To evaluate how variations in antibody structure or conjugation chemistry affect internalization and intracellular trafficking.
Preclinical development: To confirm that the selected ADC candidate exhibits consistent internalization behavior across relevant cell models.
Mechanism-of-action studies: To validate the intracellular delivery of the payload and its correlation with cytotoxic effects.
Validation Data
FACS Analysis of Antibody Internalization
FACS Analysis of Antibody Internalization

Anti-CD20 Abs and Human IgG1 isotype control were labeled with Antibody Internalization Detection Reagent (Cat.No.IGG-PZF2001). Raji cells were treated with Anti-CD20 Abs-Internalization Detection Reagent conjugate and Isotype control-Internalization Detection Reagent conjugate separately for 2 hours, then analysis by Flow cytometric. APC signal was used to evaluate the activity (Routine tested).

FACS Analysis of Antibody Internalization

Anti-Her2 Abs and Human IgG1 isotype control were labeled with Antibody Internalization Detection Reagent (Cat.No.IGG-PZF2001). SK-BR-3 cells were treated with Anti-Her2 Abs-Internalization Detection Reagent conjugate and Isotype control-Internalization Detection Reagent conjugate separately for 2 hours, then analysis by Flow cytometric. APC signal was used to evaluate the activity (Routine tested).

Fluorescence Imaging of Antibody Internalization
Fluorescence Imaging of Antibody Internalization

SK-BR-3 cells were treated with CellLights Lysosome GFP (green) for 16 hours followed by treatment with Anti-Her2 Abs-Internalization Detection Reagent conjugate and IgG1 Isotype-Internalization Detection Reagent conjugate separately for 16 hours (red), then stained with NucBlue Live ReadyProbes(blue) for 20 minutes and imaged on the EVOS M7000. A. Antibody Internalization Detection Reagent (Cat.No.IGG-PZF2001). B. IgG1 Isotype-Internalization Detection Reagent conjugate. C. Anti-Her2 Abs-Internalization Detection Reagent conjugate. D. Anti-Her2 Abs-Internalization Detection Reagent conjugate(Z-stacking).

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Resources

Webinar playback and appointment

Addressing Challenges in Antibody-Drug Conjugate Development
Here is the brief recap of the webinar:ADCs have the potential to enable 'precision medicine' with a wide market reach.Challenges in ADCs include managing Target Affinity, enhancing payload conjugation, and assessing Payload Delivery in vivo Pharmacokinetics. We can provide high-quality solutions to address these challenges.
Addressing Challenges in Antibody-Drug Conjugate Development
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Building the Perfect Antibody-based Therapeutic from Selection to Engineering and Manufacturing
In this webinar, discover how advanced techniques such as Al-driven candidate selection optimization of antibody sequences, and precise conjugation methods come together to address some of the current challenges in antibody-based therapeutics.
Addressing Challenges in Antibody-Drug Conjugate Development
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Resource download

AGLink® ADC site-specific conjugation kit: Powering your magic bullets
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Fc Receptor Proteins - Partners for Antibody Drug Development
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References

1. Beck, A., Goetsch, L., Dumontet, C. et al. Strategies and challenges for the next generation of antibody–drug conjugates. Nat Rev Drug Discov 16, 315–337 (2017). https://doi.org/10.1038/nrd.2016.268

2. Romero, D. Pembrolizum*b tunes up chemotherapy in cervical cancer. Nat Rev Clin Oncol 18, 747 (2021). https://doi.org/10.1038/s41571-021-00570-5

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