Pierce Protein AG Magnetic Beads

The Thermo Scientific Pierce Protein A/G Magnetic Beads are high-performance affinity particles for antibody purification and immunoprecipitation methods using manual or robotic magnetic separators.

These magnetic beads are coated with genetically engineered Pierce Protein A/G, a recombinant fusion protein which combines the IgG binding domains of both Protein A and Protein G. This enables capture of antibodies from a wider range of species and isotypes than either protein alone. Using our crosslinker chemistry, you can immobilize an antibody onto the magnetic particle and prevent IgG contamination in your immunoprecipitated sample. These beads can be used both manually with a magnetic stand as well as with automated platforms such as the Thermo Scientific KingFisher Instruments.

Learn more about automated magnetic separations and Thermo Scientific Automated Solutions.

Highlights:

• High capacity – nearly four times higher binding capacity than typical magnetic beads from other suppliers, allowing the use of smaller amounts per experiment
• Low non-specific binding – stable, pre-blocked beads provide clean purification products (e.g., antigen eluted in IP with antibody is devoid of contaminating proteins from complex IP matrix)
• Flexibility – convenience of IgG binding domains of both Protein A and Protein G on one bead
• Compatibility – beads are compatible with manual and automated applications (e.g., Thermo Scientific KingFisher Instruments)
• Assay consistency – magnetic beads eliminate resin loss and provide for more efficient separation of solutions than traditional IP methods that use only microcentrifuge tubes

Applications:

• IP and Co-IP experiments (see complete kit)
• Immunoprecipitation for analysis in non-reducing conditions
• Antibody purification

Product Details:

The recombinant Protein A/G that is immobilized onto the Pierce Magnetic Beads is a fusion of the IgG binding domains of both Protein A and Protein G. Protein A/G contains four Fc-binding domains from Protein A and two from Protein G, making it a convenient tool for investigating and purifying immunoglobulins. Thus, Pierce Magnetic Particles are not simply a mixed immobilization of separate Protein A and Protein G polypeptides, nor are they a mixture of Protein A magnetic beads and Protein G magnetic beads.

Diagram of Thermo Scientific Pierce Protein A/G Magnetic Beads. The magnetic particles are 1µm in diameter and are specially manufactured with two layers of magnetite and encapsulation. Recombinant Protein A/G is coupled to the bead surface.

Thermo Scientific Pierce Protein A/G Magnetic Beads isolate significantly more IgG from rabbit and mouse serum with less background than other brands of Protein A and Protein G magnetic particles. Using a KingFisher Flex with a 96 deep well plate, IgG was purified from 5mg of rabbit and mouse serum using 50µL of Pierce Protein A/G Magnetic Beads, Dynabeads* Protein A or G (Life Technologies), or PureProteome* Protein A or G Beads (Millipore). The beads were washed with Tris buffered saline containing 0.05% Tween-20 (TBST), incubated 1 hour with serum diluted in TBST, washed three times, and then eluted with 0.1M glycine, pH 2.8 for 10 minutes at room temperature. The eluates were resolved by SDS-PAGE and stained with Thermo Scientific Imperial Protein Stain. Panel A: Rabbit serum; Panel B: Mouse serum. The IgG heavy chain bands were quantified by densitometry. The values for each set of duplicate bands were averaged and expressed as a percentage of the average for the Thermo Scientific Protein A/G Beads.

The rabbit IgG Binding Capacity of Thermo Scientific Pierce Protein A/G Magnetic Beads is approx. 4-fold greater than the leading brand of Protein A and Protein G beads. Thermo Scientific Pierce Protein A/G Magnetic Beads or Dynabeads* Protein A or Protein G (Life Technologies) were added to a 96 deep-well plate (1mg beads per well). Using the Thermo Scientific KingFisher 96, the beads were washed with phosphate buffered saline containing 0.05% Tween-20 (PBST). The beads were then incubated for 1 hour with varying amounts of purified rabbit IgG (20 to 200µg). After binding, the beads were washed three times with PBST and eluted at 96°C with SDS-PAGE reducing sample buffer. Binding was calculated by subtracting the amount of protein in the flow-through from the amount of protein loaded using the Thermo Scientific BCA Protein Assay (Part No. 23225).

The Thermo Scientific Pierce Protein A/G Magnetic Beads effectively immunoprecipitate (IP) cell cycle proteins Cyclin D, Cyclin E, Cyclin B and Cdk1. U2OS (human osteosarcoma) cells were synchronized at G0 followed by growth in 20% fetal bovine serum for 4, 6 and 18 hours prior to harvest. The cells were lysed in IP Lysis/Wash Buffer, and 0.75mg of lysate (per sample) was incubated with anti-Cyclin D (rabbit polyclonal), anti-Cyclin E (mouse IgG1), anti-Cyclin B (mouse IgG1), or anti-Cdk1 (rabbit polyclonal) antibodies overnight at 4°C. The Pierce Protein A/G Magnetic Beads were added (50µL each) to a 96 deep-well plate and immunoprecipitations were performed using the KingFisher Flex. Eluted sample volumes of 5µL, 10µL and 20µL were resolved by SDS-PAGE and analyzed by Western blot.

Related Products:

Pierce Protein A/G Magnetic IP/Co-IP Kit
Other Pierce Magnetic Beads
Pierce Protein A/G Agarose Beads
Thermo Scientific KingFisher Magnetic Particle Processors