At the end of a cycle of immunodetection, it is possible to strip the membrane with stripping buffer and to carry out subsequent cycles incubating with different primary antibodies (see Note 21).
The procedure for the stripping we use is as follows:
1. Incubate the membrane in stripping buffer at 70°C for 30 min, with occasional shaking.
2. Wash the nitrocellulose 2 x 10 min in large volumes of washing solution (Subheading 2.2., step 12) at room temperature.
3. Block the membrane and perform immunodetection as described in
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