Fab-pHast products bind to your primary antibody via a secondary antibody or Streptavidin cross-linked to a pH-dependent fluorescent reporter. This fluorescent reporter increases intensity as the pH of its surroundings becomes more acidic, as evident when exposed to the environment inside a cell.
SLC46A3 as a Potential Predictive Biomarker for Antibody-Drug Conjugates Bearing Noncleavable Linked Maytansinoid and Pyrrolobenzodiazepine Warheads.
Kinneer K, et al. Clin Cancer Res, 24 (24):6570-6582, 2018.
Objective: To develop biomarkers to uncover the underlying mechanism of resistance by certain cell lines for ADCs.
Summary: Loss of SLC46A3 expression was found to be a mechanism of innate and acquired resistance to ADCs bearing DM1 and SG3376.
Dose: For Lysosomal trafficking, ADCs were labeled with Fab-pHast human (Cat. #PH-01). Cells were incubated with 3 mg/mL of labeled ADCs at 37°C for desired time points and fluorescence quantified by flow cytometry.
pHast Ab Internalization Assay
Parental HEK-293 cells, and HEK-293 cells transfected with the p75 receptor, were plated in a 96-well plate overnight. Titrated 192-IgG antibody (Cat. #AB-N43) was incubated at RT with 50 nM of Fab-pHast Mouse (Cat. PH-02) for 20 min prior to addition to cells. Plates were incubated overnight to allow maximum internalization, but a few hours is sufficient for detection.
Please let us know if we can answer any questions about this promotion or any of our products.