Thermo Scientific Pierce Anti-HA Magnetic Beads are affinity particles for immunoprecipitation of recombinant HA-tagged proteins expressed in bacterial or mammalian cells or in vitro systems, using manual or robotic magnetic separators.
The blocked magnetic bead surface is coated with anti-HA antibody, a highly specific mouse IgG1 monoclonal antibody that recognizes the HA-epitope tag (YPYDVPDYA) derived from the human influenza hemagglutinin (HA) protein. The Pierce Anti-HA Magnetic Beads can be used manually with a magnetic stand, as well as with automated platforms, such as the Thermo Scientific KingFisher Instruments, for high-throughput workflows.
- Specific – highly specific anti-HA monoclonal antibody (clone 2-2.2.14) enables high yield and high purity immunoprecipitation
- Convenient and fast – product instructions provide an easy-to-follow, optimized protocol for immunoprecipitation in approximately one hour
- Low non-specific binding – stable, pre-blocked beads and specific antibody minimize off-target binding for HA-tag IP or co-IP experiments
- Versatile – beads are compatible with manual and automated workflows (e.g., Thermo Scientific KingFisher Instruments)
Pierce Anti-HA Magnetic Beads are convenient for the immunoprecipitation (IP) of recombinant HA-tagged proteins and the co-immunoprecipitation (co-IP) of their interacting proteins. The beads are incubated with a cell lysate containing HA-tagged protein and the fusion protein is captured. The beads are subsequently washed and then the target proteins are eluted using 0.1M glycine, pH 2.0, 50mM NaOH, or SDS-PAGE sample buffer. If gentler elution conditions are desired, 2mg/mL Pierce HA peptide can be used. The protocol has been optimized for each of these conditions. Anti-HA antibody can be used to detect HA-tagged protein by Western blot analysis.
Characteristics of Thermo Scientific Pierce Anti-HA Magnetic Beads.
||High-affinity mouse IgG1 monoclonal antibody (clone 2-2.2.14) covalently coupled to the surface of blocked magnetic beads
||Superparamagnetic (no magnetic memory)
||≥10μg of GST-ERK-HA (70kDa fusion protein)/mg of beads;
≥100μg of GST-ERK-HA (70kDa fusion protein)/mg of bead suspension
|Better immunoprecipitation results with Thermo Scientific Pierce Anti-HA Magnetic Beads. Using a KingFisher Flex Instrument with 96 deep-well plates, 25μL each of Pierce Anti-HA Magnetic Beads, Anti-HA-tag Magnetic Beads (MBL International Corp., Supplier 1) and SPHERO™ Rabbit Anti-HA Magnetic Beads (Spherotech Inc., Supplier 2) were used to immunoprecipitate GST-PI3K-SH2-HA from 50μg of E. coli lysate in duplicate. Captured protein was eluted with 0.1M glycine, pH 2.0, and then resolved by SDS-PAGE and analyzed by Western blot for the HA-tagged protein.
||Significantly better binding capacity with Thermo Scientific Pierce Anti-HA Magnetic Beads. Binding capacity was determined manually using a magnetic stand. HA-tagged protein (HA-ERK-GST, 400μg in PBS) was incubated with 100μL each of Pierce Anti-HA Magnetic Beads and Anti-HA-tag Magnetic Beads from MBL International for 1 hour at room temperature. Bound HA-tagged protein was measured using the Thermo Scientific Pierce BCA Assay by subtracting the amount of protein in the flow-through from the amount loaded onto the beads. The Pierce beads bound four-fold more protein than the equivalent amount of MBL beads.
HA-Tag Magnetic IP/Co-IP Kit (magnetic)
HA-Tag IP/Co-IP Kit (agarose)
Anti-HA Agarose and Peptide
Anti-HA Antibody and other Anti-Tag Antibodies
KingFisher Magnetic Particle Processors
Cell Lysis and Protein Extraction Reagents