Fab-pHast mouse is one of our fastest tools for quantitative testing of your primary antibody’s specificity, binding, and internalization, providing results in 1 day. Fab-pHast mouse binds to your primary mouse antibody via a secondary antibody cross-linked to a pH-dependent fluorescent reporter. This fluorescent reporter will increase 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 antibody candidates.
Fab-pHast mouse generates quantitative testing of the specific, binding, and internalization of your primary mouse antibody, with results in 1-day. This secondary conjugate is used to evaluate the potential of a primary antibody to internalize. Making a 1.5 ml “Fab-pHast spiked solution” will use about 43 micrograms of Fab-pHast. This is assuming a 500-nM spiked solution and performing an assay on a 96-well plate, which is the suggested amount on the protocol. This is enough to test 1 antibody in sextuplicate (6 data points) at 8 different concentrations of antibody.
Other pHast Conjugates:
Also available: Streptavidin-pHast (PH-03). Streptavidin-pHast binds to your biotinylated protein and contains a pH-dependent fluorescent reporter (pHast).