Pierce™ Protein A/G Magnetic Beads, 5 mL - FAQs

View additional product information for Pierce™ Protein A/G Magnetic Beads - FAQs (88803, 88802)

8 product FAQs found

Can I reuse Pierce Protein A/G Magnetic Beads (Cat. No. 88802, 88803)?

Pierce Protein A/G Magnetic Beads are intended for single use only. Therefore, we do not recommend reusing them.

What advantage do Pierce Magnetic Agarose beads have over Dynabeads or Pierce Magnetic Beads?

Pierce Magentic Agarose Beads have a much higher binding capacity.

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.

Can I centrifuge the Thermo Scientific Protein A/G Magnetic Beads?

No, centrifuging causes the formation of irreversible aggregates which greatly reduces binding capacity.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

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.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

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.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

How can I prevent bead aggregation with Thermo Scientific Protein A/G 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 0.05% non-ionic (e.g., Tween-20 Detergent) or zwitterionic (e.g., CHAPS) detergent in the binding buffer and mix the beads during incubation.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

Will the Thermo Scientific Protein A/G Magnetic Beads bind to goat IgG?

Yes, Protein A/G magnetic beads will bind to goat IgG as well as several other species of IgG such as rabbit, mouse, etc. For details of species as well as different immunoglobulin isotypes that will bind to Protein A/G, please look at this Tech Tip (http://tools.thermofisher.com/content/sfs/brochures/TR0034-Ab-binding-proteins.pdf).

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

What are the differences between Protein A, Protein G, Protein A/G, and Protein L Ig-binding proteins?

Protein A, Protein G, and Protein A/G bind almost exclusively to the IgG class of antibodies, but their binding properties differ among species and subclasses of IgG. Protein L binds in the variable fragment of some kappa light chains and can react with any immunoglobulin, not just IgG, as long as the correct kappa light chains are present. Protein L does not bind lambda light chains and certain kappa chains of different species.

-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 (e.g., IgG1 vs. IgG2)
-Protein A/G is a recombinant fusion protein that includes the IgG-binding domains of both Protein A and Protein G. Therefore, Protein A/G is ideal for binding the broadest range of IgG subclasses from rabbit, mouse, human, and other mammalian samples.
-Protein L binds to certain immunoglobulin kappa light chains. Because kappa light chains occur in members of all classes of immunoglobulin (i.e., IgG, IgM, IgA, IgE and IgD), Protein L can purify these different classes of antibody. However, only those antibodies within each class that possess the appropriate kappa light chains will bind. Generally, empirical testing is required to determine if Protein L is effective for purifying a particular antibody. It binds only Vk1 in mouse and VkI, VkIII and VkIV in human.
Read more about the general characteristics of Ig-binding proteins (https://www.thermofisher.com/us/en/home/life-science/antibodies/antibody-purification-kits-reagents.html) and (https://assets.thermofisher.com/TFS-Assets/LSG/Application-Notes/TR0034-Ab-binding-proteins.pdf).

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.