Improved Multiplexing with Simple Western using a 20X Rabbit HRP ConjugateTechnical Notes
Technical Notes Summary
Multiplexing two targets in one capillary with Simple Western doubles the number of data points per sample and increases the data quantitation accuracy when you normalize to biological loading control or system control. When you're ready to start multiplexing, first optimize the primary antibodies separately to find the saturating dilution for each. Then combine them in Antibody Diluent 2 to detect the two targets in the same capillary. If the primary antibodies are raised in two different host species, you'll also need to combine two different anti-species secondary antibodies. But mixing Ready-Use (RTU) secondary antibodies will dilute them and cause the signal to decrease or become more variable because they're no longer saturating. We've got the situation under control with the 20X Anti-Rabbit HRP Conjugate.
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