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Product Details
Application
Antibody Internalization Detection Reagent (Cat. No. IGG-PZF2001) utilizes an anti-human Fc antibody conjugated to a pH-sensitive fluorescent dye, specially engineered for acid-triggered signal amplification. This reagent specifically labels the Fc region of human IgG antibodies to form a stable complex, enabling real-time monitoring of antibody endocytosis in live cells. The pH-sensitive dye cannot only significantly enhance the fluorescence signal in an acidic environment but also minimize background noise effectively.
Features
- High signal-to-noise ratio - Strong fluorescence with minimal background
- Rapid fluorescent labeling - Complete in 10 minutes
- pH sensitive - Bright emission in acidic milieu
- Fc-Specific targeting - Preserves Fab antigen-binding integrity
Specificity
Specifically recognizes the human antibody Fc portion.
Detection Wavelength
Excitation Wavelength: 643 nm
Emission Wavelength: 660 nmStorage
For long term storage, the product should be stored at lyophilized state at -20°C or lower.
Please protect from light and avoid repeated freeze-thaw cycles.
This product is stable after storage at:
- -20°C to -70°C for 24 months in lyophilized state;
- -20°C to -70°C for 12 months after reconstitution.
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Performance Data
Bioactivity-FACS
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 (QC tested).
Protocol
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 (Routinely tested).
Protocol
FACS Data
In the assessment of ADC efficacy using the Antibody Internalization Detection Reagent (ACROBIOsystems, Cat. No. IGG-PZF2001) with HER2-positive SK-BR-3 cells and HER2-negative MFI-MDA-MB-468 as target cells; (A) Prolonged co-incubation of HER2-targeting antibodies with HER2-positive SK-BR-3 cells enhanced antibody internalization. (B) whereas no internalization was observed in HER2-negative MFI-MDA-MB-468 cells.
Protocol
In the assessment of ADC efficacy using the Antibody Internalization Detection Reagent (ACROBIOsystems, Cat. No. IGG-PZF2001) with HER2-positive SK-BR-3 cells as target cells; (C) The trastuzumab-based ADC effectively internalizes relative to its naked antibody counterpart at concentrations ≥0.1 μg/ml, demonstrating dose-dependent enhancement with comparable efficacy between the two. (D) Disitamab-based ADC effectively internalizes relative to its naked antibody counterpart at concentrations ≥0.1 μg/ml, demonstrating dose-dependent enhancement, with no statistically significant difference observed between the two.
Protocol
Fluorescence Imaging
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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