Western blot Protocol
1. Block membrane with 5% non-fat milk in PBS-T for 1 hour at room temperature or longer at 4C.
2. Incubate membrane with IgY antibodies at dilution of 1: 3,000 with 1% milk in PBS-T at R.T. for 1 h.
3. Rinse 3 times with PBS-T, then wash membrane with PBS-T, 5 min each, total of 3 times. 4. Incubate with 2nd antibody (goat-anti-IgY/Fc-HRP) at dilution 1:10,000 for ECL (with 1%milk PBS-T) at R.T. for 1h.
5. Rinse 3 times with PBS-T, then wash with PBS-T, 5 min each with shaking, total of 3 times.
6. Perform ECL detection of signal using Pierce ECL kit.