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Thermo Scientific™ Pierce™ Protein A Spin Plate for IgG Screening

Catalog No. PI45202
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Catalog No. PI45202 Supplier Thermo Scientific™ Supplier No. 45202
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Affinity purify antibodies (IgG) with densely immobilized Protein A offered in bottles, spin columns, filter plates and complete purification kits.

Thermo Scientific™ Pierce Protein A Plus Agarose is a versatile, high-performance affinity resin for antibody purification that is available as bottled agarose beads, pre-packed spin columns, 96-well filter plates and complete IgG purification kits.

Pierce Protein A Plus Agarose consists of purified native Protein A that has been covalently immobilized at high density onto high-quality crosslinked 6% beaded agarose (CL-6B). Among the many available forms of immobilized Protein A affinity resin, this premiere variety provides the most versatile combination of chromatographic features for high yield and high purity purification of whole IgG from mammalian serum samples. The agarose beads have physical and chemical properties suitable for many affinity purification systems. Accordingly, Pierce Protein A Plus Agarose is offered in several convenient package formats, including bottled resin slurries, three sizes of spin columns, complete purification kits, two sizes of FPLC-ready chromatography cartridges, and 96-well filter plates.

Protein A is a bacteria-derived protein that binds with high affinity and specificity to the Fc portion of antibodies, especially those of the IgG class. Compared to its alternative (Protein G), Protein A provides the highest binding capacity for subclasses of IgG from rabbits, pigs, dogs and cats. Protein A also has higher binding capacity for human IgG, except for IgG3, which it binds weakly. Protein A binds weakly to mouse IgG1 and is not recommended for most applications with that antibody isotype.

Highlights:

  • Native Protein A – immobilized Protein A is ideal for polyclonal IgG purification from human, rabbit, pig, dog or cat serum
  • Agarose resin – support is crosslinked 6% beaded agarose (CL-6B), the most popular resin for protein affinity purification methods
  • Inert and stable – superior manufacturing method immobilizes Protein A by charge-free, leach-resistant covalent bonds, resulting in low nonspecific binding and enabling multiple uses without decline in yield
  • High capacity – “Plus” variety of Pierce Protein A Agarose has a dense load of immobilized Protein A, providing a binding capacity greater than 34mg human IgG/mL resin (approx. 16 to 17mg mouse IgG/mL resin)
  • Multiple formats – choose from bottled resin slurries, centrifuge-ready columns, complete purification kits, two sizes of FPLC-ready chromatography cartridges, and 96-well filter plates

Properties of crosslinked 6% beaded agarose (CL-6B):

  • Support pH Stability: 2 to 14 (short term); 3 to 13 (long term)
  • Average Particle Size: 45 to 165 microns
  • Exclusion Limit: 10,000 to 4,000,000 daltons
  • Maximum Volumetric Flow Rate: approx. 1mL/minute (for 1cm diameter column)
  • Maximum Linear Velocity: 30cm per hour
  • Maximum Pressure: less than 25psi (1.5 bar), defined as the maximum pressure drop across a column that the resin can withstand (Note: The indicated gauge pressure of a liquid chromatography apparatus may be measuring the total system pressure rather than the pressure drop across the column.)

Available Formats:

  • Resin Slurries: 1mL, 5mL, 25mL bottles
  • Spin Columns: 0.2mL (microcentrifuge), 1mL, 5mL columns
  • Purification Kits: complete kits in all three column sizes
  • Chromatography Cartridges - compatible with automated liquid chromatography systems and with syringe processing
  • 96-well Spin Plates: pre-dispensed 96-well filter plates, compatible with centrifugation and vacuum manifold systems for manual or automated purification. The plates enable fast, consistent well-to-well and plate-to-plate reproducibility for small-scale, high-throughput separations
  • Bulk and Custom: let us know your needs for bulk Pierce Protein A Plus Agarose

Includes:

Purification kits contain Protein A Plus Spin Columns, binding buffer, elution buffer and neutralization buffer

Specifications

Product Type Spin Plate
Product Line Pierce
Column Type Agarose Resin, Affinity
Format 96-well Filter Plate
Stationary Phase Protein A
Purification Target IgG Antibodies
Description Pierce Protein A Spin Plate for IgG Screening
Content And Storage Upon receipt store at 4°C
Quantity 1 Plate
Can I use the Protein A or Protein G Antibody Screening Spin Plate to purify chicken IgY?

No. Neither Protein A nor Protein G binds IgY. Thermo Scientific Thiophilic Adsorbent (Cat. Nos. 20500 and 44916) purifies IgY from serum. The Thermo Scientific Chicken IgY Purification Kits (Cat. Nos. 44918 and 44922) purify IgY from yolks.

Can I reuse the Protein A and Protein G Antibody Screening Spin Plates?

Yes. To regenerate the resins, wash wells three times with elution buffer and three times with 0.02% sodium azide in ultrapure water as detailed in the instructions.

Can I use just a portion of the Antibody Screening Spin Plate to process a few samples?

No. The Protein A or Protein G 96-well Spin Plates have bottom and top seals. Removing the seals will cause the buffers to leak into the wash/collection plate. It is not possible to remove only a portion of the seal and continue with the purification process, and therefore, process 96 samples each time.

What are the binding capacities of the Antibody Screening Spin Plates?

The Protein A Spin Plate binds 0.6 mg of human or 0.3-0.4 mg of mouse IgG/well (based on > 12 mg of human IgG or 6-8 mg of mouse IgG/mL of settled resin). The Protein G Spin Plate will bind > 0.5 mg human IgG/well (based on 11-15 mg of human IgG/mL of settled resin).

What is the difference between Protein A and Protein G binding capabilities?

Protein A and Protein G bind almost exclusively with the IgG class of antibodies, but their binding properties differ among species and subclasses of IgG. Protein A is generally preferred for rabbit, pig, dog and cat IgG. Protein G has better binding capacity for a broader range of mouse and human IgG subclasses (IgG1, IgG2, etc.), and it binds rat IgG2a and IgG2b, which do not bind to Protein A. For a comprehensive list of the binding characteristics of Protein A and Protein G, see Tech Tip #34 (https://assets.thermofisher.com/TFS-Assets/LSG/Application-Notes/TR0034-Ab-binding-proteins.pdf).

What is included with the Antibody Screening Spin Plates?

Each product includes one 96-well Spin Plate and three Wash/Collection plates (buffers are not included). Each of the 96 wells contains 50 µl of agarose resin (200 µl of 25% slurry) with the immobilized protein and can process 10-100 µl of sample.

In what applications can I use the Antibody Screening Spin Plates?

Protein A (Cat. No. 45202) and Protein G (Cat. No. 45204) Spin Plates for IgG Screening are for rapid, small scale, affinity-purification of polyclonal and monoclonal antibodies from serum, ascites and culture supernatants.


For Research Use Only. Not for use in diagnostic procedures.