Rules for Multiple Label Experiments

In previous experiments with single 1° antibodies (Chapter 10), the 2° antibody control showed that 2° antibody bound correctly and that its concentration was appropriate. With multiple P antibody experiments, the possibility exists that the 2° antibodies will bind unexpectedly in unforeseen ways. In the controls with multiple antibodies, an additional set of control conditions must be done each time the experiment is performed to insure that only the correct pair of 1° and 2° antibodies will bind. 

The first set of controls are no 1° for each of the P antibodies (Table 11.2). This should show that the labeling for the 1° antibody was omitted and is not seen. The second control is to use neither of the 1° antibodies and eliminate all of the labeling. 

The important rules listed here summarize ideas from this chapter and the preceding chapters. 

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