Antibody Internalization Detection Reagent

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).

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).

Trontinemab and Human IgG1 isotype control were labeled with Antibody Internalization Detection Reagent (Cat. No. IGG-PZF2001). HEK293/Human Transferrin R Stable Cell Line were treated with Trontinemab-Internalization Detection Reagent conjugate and Isotype control-Internalization Detection Reagent conjugate separately for 4 hours, then analyzed by Flow cytometric. APC signal was used to evaluate the activity (Routinely tested).

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.

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.

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).