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  • Protocol for Anti-SNAP-tag® Antibody (Polyclonal) (P9310)

    Protocol

    1. Perform SDS-PAGE and protein transfer according to standard Western blotting methods.
    2. After protein transfer, wash membrane with 25ml of 1X TBS for 5 minutes at room temperature.
    3. Incubate membrane in 25ml of Blocking Buffer for 1 hour at room temperature.
    4. Wash membrane 3 times for 5 minutes each with 15ml of TBST Wash Buffer.
    5. Incubate membrane with Anti-SNAP-tag® Antibody (NEB #P9310S) at a 1:1000 dilution in 20ml of Blocking Buffer with gentle rocking overnight at 4ºC or for 1 hour at room temperature.
    6. Wash membrane 3 times for 5 minutes each with 15ml of TBST Wash Buffer.
    7. Incubate membrane with a commercially available secondary antibody (e.g. Anti-Rabbit IgG HRP-linked antibody) in 20ml of Blocking Buffer with gentle rocking for 1 hour at room temperature. Use the working dilution suggested by the manufacturer of the secondary antibody.
    8. Wash membrane 3 times for 5 minutes each with 15ml of TBST Wash Buffer.
    9. Proceed to chemiluminsecent detection of protein bands with a commercially available detection reagent.