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应用（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.
优势特色（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
特异性（Specificity）
Specifically recognizes the human antibody Fc portion.
检测波长（Detection Wavelength）
Excitation Wavelength: 643 nm
Emission Wavelength: 660 nm存储（Storage）
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.
质量管理控制体系（QMS）
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数据展示
活性（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
流式数据 -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
荧光成像（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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