Fab-pHast human 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 human binds to your primary human 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 human generates quantitative testing of the specific, binding, and internalization of your primary human antibody, with results in 1-day. This secondary conjugate is used to evaluate the potential of a primary antibody to internalize. Fab-pHast reacts with whole molecule IgG (heavy and light chains).
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:
View a table of targets and recommended pHast conjugates for testing your primary antibody specificity.
Also available: Streptavidin-pHast (PH-03). Streptavidin-pHast binds to your biotinylated protein and contains a pH-dependent fluorescent reporter (pHast).