用法 :
•Do not centrifuge, dry or freeze Anti-HA Magnetic Beads. Centrifuging, drying or freezing will 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) or zwitterionic (e.g., sc-29088, CHAPS) detergent in the binding and wash buffers and to mix the beads during incubation.
• For best results, determine optimal conditions for expression of HA-tagged fusion protein before attempting immunoprecipitation.
• To minimize protein degradation, include protease inhibitors (such as: UltraCruz® Protease Inhibitor Cocktail Tablet or UltraCruz® Protease Inhibitor Cocktail Tablet, EDTA-free) in the preparation of cell lysates.
• Binding capacity and elution recovery will vary depending on the HA-fusion protein and the method of elution.
• 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 HA 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 HA-tagged protein; however, the stringency of the buffer will cause the HA antibody to leach from the beads.
• If a gentle elution of HA-tagged protein is desired, a competitive elution can be performed using 2mg/mL HA Peptide.
• Anti-HA Magnetic Beads are compatible with immunoprecipitation and analyses by Western blot.
• Do not use cell lysate containing dithiothreitol (DTT). DTT may cause the HA antibody to leach from the beads.