The UltraTag Hapten Labeling Kits efficiently conjugate 50 µg of antibody in 50 µL with one of four available hapten tags in less than an hour, with ~100% tagging, >80% recovery, and with less than two-fold dilution.
Then, the tagged antibody can be used in any immunoassay and subsequently detected by the corresponding anti-hapten antibody. Up to four antibodies tagged with the four different haptens can be used together and detected
independently. Does not interfere with other methods, such as conventional detection, and is compatible with biotin or digoxigenin detection, to allow independent analysis of up to six antigens.
Independently detect multiple monoclonal and/or polyclonal primary antibodies from the same or different species in one experiment
2, 3, or 4 color immunofluorescence on tissues, cells, spheroids, or organoids
2 or 3 chromogen immunohistochemistry on cell spreads or tissue sections
Use in other immunoassays, e.g., EliSpot, flow cytometry, ELISA
Standard, two step staining protocols are used, applying the hapten-labeled primary antibodies and then the
fluor-labeledanti-hapten secondary antibodies as cocktails
Four-hapten/fluor-labeledanti-hapten antibody pairs available for multiplexing
Overcomes primary antibody species limitations
Overcomes species-on-species cross-reactivity
Sensitivity for each primary antibody remains nearly the same as for conventional anti-species secondary antibody detection
Allows mixing-and-matching of fluors or enzymes for any antibody
Figure 1. Scheme of UltraTag mxIF multiplex immune-fluorescence staining.
Figure 2. Multiplex image of tonsil tissue stained with hapten-labeled CD8 (green), CD4 (purple), PD-1 (red), and FoxP3 (yellow) prepared using the Cell IDx UltraTag Hapten Labeling Kit.