The pHast technology is one of our fastest tools for quantitative testing of your material’s specificity, binding, and internalization, providing results in 1 day.  pHast products bind to your material 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. A successful assay will provide an EC50 by way of a fluorescence detecting plate reader, illuminating your lead candidates.

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 was incubated at RT with 50 nM of Fab-pHast Mouse for 20 min prior to addition to cells. Plates were incubated overnight to allow maximum internalization, but a few hours is sufficient for detection.