How do I reduce non-specific binding to Thermo Scientific Protein A/G Magnetic Beads?
Adding 50-350 mM of NaCl to the Binding/Wash and Elution Buffers can help reduce non-specific bands. Also, use a low-pH elution for single-use applications. The optimal time for low-pH elution is 10 minutes; exceeding 10 minutes may result in non-specific binding and yield reduction.
When should I avoid boiling the Thermo Scientific Protein A/G Magnetic Beads?
Avoid bead-boil elutions when using rabbit antibodies (primary or secondary) in downstream Western blot applications. Instead, perform elution in SDS-PAGE sample buffer at room temperature. For all other antibody species, boiling the beads in SDS-PAGE sample buffer is acceptable for single-use applications. Boiling could cause bead aggregation and loss of binding activity.