用法 :
• Do not centrifuge, dry or freeze Anti-c-Myc Magnetic Beads, as this can cause the beads to aggregate and lose binding activity. To ensure good dispersal of beads for optimal antibody binding, it is important to include 0.025% to 0.1% non-ionic (e.g., Tween-20 Detergent: sc-29113, ) or zwitterionic (e.g., CHAPS: sc-29088) detergent in the binding and wash buffers and to mix the beads during incubation.
• For best results, determine optimal conditions for expression of c-Myc-tagged fusion protein before attempting immunoprecipitation.
• To minimize protein degradation when preparing cell lysates, include protease inhibitors such as:
• UltraCruz® Protease Inhibitor Cocktail Tablet: sc-29130
• UltraCruz® Protease Inhibitor Cocktail Tablet, EDTA-free: sc-29131
• Binding capacity and elution recovery will vary depending on the c-Myc-fusion protein and the elution method.
• A low-pH elution may be used for single-use applications. Optimal incubation time for low-pH elution is 5-10 minutes; exceeding 10 minutes may result in nonspecific binding and yield reduction. The c-Myc antibody will not leach from the beads when eluting with the recommended acidic elution buffer (0.1M glycine, pH 2.0).
• Basic elution buffer (e.g., 50mM NaOH) may be used to elute c-Myc-tagged protein; however, the stringency of the buffer will cause the c-Myc antibody to leach from the beads.
• If a gentle elution of c-Myc-tagged protein is desired, a competitive elution can be performed using 0.5mg/mL of c-Myc Peptide.
• Anti-c-Myc Magnetic Beads are compatible with IP and Western blot analysis.
• Do not use cell lysate containing dithiothreitol (DTT). DTT may cause the c-Myc antibody to leach from the beads.