Immunoprecipitation Magnetic Beads

Dynabeads magnetic beads offer the best balance of capacity/yield, reproducibility, purity, and cost for smaller-scale isolation of specific proteins (e.g., immunoprecipitation, IP) and protein complexes (co-immunoprecipitation, co-IP). Dynabeads is continually shown to be the fastest growing method for Immunoprecipitation in around the world.

Buy nowSee Immunoprecipitation app notes

Dynabeads features

  • Low background—little to no nonspecific binding and no preclearing required for high-purity protein yields.
  • Highly reproducible—uniform beads ensure the most consistent results.
  • Highly sensitive—most-cited method for sensitive applications, such as ChIP and IP, of low- abundance proteins.
  • Fast and easy—<40 min protocol with no centrifugation or preclearing steps.
  • Antibody savings—all binding occurs on the smooth outer surface of the beads, which conserves precious antibodies and supports a cost-efficient solution per sample.
  • Flexible—products for IP, Co-IP, pull-down, and ChIP assays; ideal for manual and automated protocols.
  • Automation of IP protocol—learn how the Thermo Scientific KingFisher platform now supports immunoprecipitation automation.

How to perform Immunoprecipitation using Dynabeads

Step 1: Binding of the antibody to the beads

Binding of the antibody to the beads. Dynabeads magnetic beads are precoated with protein A, protein G, anti-mouse IgG, or anti-rabbit IgG antibodies and due to the rapid kinetics will bind the added antibody in only 10 minutes. Biotinylated antibodies can also be used in combination with streptavidin-coupled Dynabeads. Wash the beads once on the magnet to remove of unbound antibodies.

Step 2: Add sample to the beads

Add sample to the beads. Add the protein-containing sample to the washed beads with bound antibody and incubate for another 10 minutes to bind the target protein of interest. If the protein is of low abundance or low affinity, the incubation can be increased to 1 hour or an overnight incubation; but in general, a 10-minute incubation is sufficient.

Step 3: Wash the protein-bound beads

Wash the protein-bound beads. To ensure high purity of the bound target protein, the beads should be washed 2–4 times with buffer on the magnet to remove all unbound proteins. The efficient isolation process ensures a high signal-to-noise ratio.

Step 4: Elute the protein

Elute the protein. If required, the protein can be eluted off the beads by using either a mild elution procedure or a denaturing elution procedure (for details, see elution description below).

Process and Benefits of IP with Magnetic Dynabeads

Protein elution properties

Protein elution properties
Varying degrees of elution with Dynabeads

The recombinant protein A and/or protein G on the beads contains no albumin binding sites, thus avoiding co-purification of contaminating proteins. Binding of antibodies to the beads in solution is an equilibrium reaction. To capture as many antibodies as possible, the reaction volume should be kept low to maintain high concentrations of beads and antibody. There is no need for pre-treating the sample (even viscous samples), hence no dilution of sample or beads should be done. If the antibody concentration in the sample is suspected to be very low, the amount of beads can be increased.

As an alternative to eluting antibody from the beads, the Dynabeads may be re-suspended in a Na-phosphate buffer. To prevent co-elution, it is possible to cross-link the antibody to protein A/G on the beads prior to immunoprecipitation. This is not necessary for downstream SDS-PAGE followed by autoradiography or western blotting, and optional for silver or coomassie dye staining.

Find the right immunoprecipitation product for your experiment

Choose this if you have an antibody that recognizes your protein

Your choice of antibody binding products depends on the downstream assay.

Antibody binding is the most common method and is used when your target antibody can be bound directly to the beads or indirectly to a pre-coated ligand on the magnetic beads.

Antibody-binding product selection guide

  Protein A, G, A/G Secondary antibodies
(anti-mouse, anti-rabbit)
Surface-activated beads (epoxy)*
Products

Beads only:

Kits:

Beads only:

Kits:

Binding properties Non-covalent antibody binding Non-covalent antibody binding Covalent antibody binding
Antibody co-eluted off the beads Yes Yes No
Type of ligand Different ligands bind different species and antibody subclasses with different specificity Anti-mouse binds mouse IgG1, IgG2a, IgG2b; anti-rabbit binds all rabbit IgGs All antibodies
Mass spec compatible Yes (Protein A/G);
Not validated (Protein A, G)
No Yes
Nonspecific binding Low Low Ultra-low
Antibody coupling time & temperature (RT=room temp.)
  • Protein A: 10 min, RT
  • Protein G: 10–40 min, RT
  • Protein A/G: 60 min, RT
>30 min, 2–8°C 16–24 hr, 37°C

* See more choices in surface-activated Dynabeads for the binding and capture of additional targets.
† Crosslinking can be performed to avoid co-elution of the antibody, but this can decrease the yield of the target antigen.
‡ Learn which antibody-binding proteins are best for your IP antibody.

See ordering information


Choose this if you have a biotinylated antibody (or ligand) that recognizes your protein

Main advantages for using a biotinylated antibody with streptavidin-coated beads for IP include:

  • If you have a sample rich in soluble IgGs
  • If you have a recombinant antibody lacking Fc regions
  • If you already have streptavidin-coated Dynabeads in the lab
  • If you need a bead compatible with mass spectrometry (secondary-coated and epoxy-coated Dynabeads are also compatible with mass spectrometry)

To allow flexibility, four different types of streptavidin-coupled Dynabeads are available. View the Dynabeads streptavidin selection guide to see the different characteristics.

Over the past 25 years, streptavidin-coupled Dynabeads in combination with a biotinylated ligand have been widely used and are cited in thousands of papers for diverse manual and automated applications. The most widely used application for Streptavidin Dynabeads is for purification of nucleic acids and next generation sequencing (NGS) workflows, but they are also used for immunoprecipitation (IP).

See ordering information


Choose this if you have a recombinant (fusion tagged) protein

Popular fusion tags for recombinant protein expression include:

  • His tag—consists of a string of six to nine histidine residues; primarily used for purification via immobilized metal affinity chromatography (IMAC)
  • GST tag—consists of glutathione S-transferase (GST), a complete 211 amino acid protein (26 kDa); primarily used for purification via glutathione agarose resin
  • HA tag—consists of a peptide (YPYDVPDYA) derived from the human influenza hemagglutinin (HA) protein; immobilized anti-HA antibodies are used to immunoprecipitate HA-tagged recombinant proteins
  • c-Myc tag—consists of a peptide (EQKLISEEDL) derived from the human c-myc oncogene (p62 c-myc); immobilized anti-c-Myc antibodies are used to immunoprecipitate c-Myc-tagged recombinant proteins.
  • FLAG tag—consists of a peptide (DYKDDDDK) which is recognized by an immobilized high-affinity rat monoclonal antibody (clone L5); primarily used for the isolation of protein complexes with multiple subunits, because the mild purification process tends not to disrupt these interactions

His and GST are not true epitope tags, because they are not usually purified or detected via specific antibodies. By contrast, HA and c-Myc tags are true epitope tags, because their only means of purification or detection is via specific antibodies. Epitope tags are seldom used for bulk purification but are most often used for immunoprecipitation or co-immunoprecipitation (IP, co-IP). Nevertheless, all four of these tag systems can be used for either purification or pull-down applications.

If you’ve got a biotinylated antibody that recognizes the protein you wish to capture, you can create your own affinity product with one of these products.

See ordering information

Benchmarking data: Dynabeads vs. other IP solutions

Dynabeads continually outperform other methods and magnetic bead solutions for IP. Still using Sepharose or agarose bead slurry. See some of the most common myths about immunoprecipitation.

Dynabeads compared to resin-based solutions

Figure 1. Shorter protocol time and better yields with Dynabeads magnetic beads. Equal amounts of sample material with respect to Ab content and cell lysate volume were used for all IP protocols according to the manufacturer’s protocol. For the Dynabeads-based method, all the antibodies on the bead surface are accessible for optimal, highly reproducible antigen binding.

Dynabeads compared to magnetic solutions

Figure 2. The magnetic bead you choose will affect your results. Dynabeads magnetic beads have a defined surface to carry out the necessary binding, with no inner surface to trap any unwanted proteins. (A) Dynabeads products are the most uniform, monodisperse superparamagnetic beads, manufactured with highly controlled product qualities to enable a high degree of reproducibility. (B–D) Magnetic particles from alternative suppliers have variable shapes and sizes that trap impurities, resulting in lower reproducibility and increased nonspecific binding.

Figure 3. Benchmarking data shows that Dynabeads magnetic products have the best performance when taking into account the combination of highest yield and lowest nonspecific binding. Our magnetic beads also have the fastest protocol, helping improve your lab life by removing unnecessary steps.

Automate immunoprecipitation with KingFisher instruments

Complete protein immunoprecipitation in 40 min with robust and consistent results using Dynabeads on KingFisher instruments.

See how Dynabeads and KingFisher can shorten Immunoprecipitation to 40 min, with only 15 min “hands on” time

KingFisher instruments

Dynabeads features

  • Low background—little to no nonspecific binding and no preclearing required for high-purity protein yields.
  • Highly reproducible—uniform beads ensure the most consistent results.
  • Highly sensitive—most-cited method for sensitive applications, such as ChIP and IP, of low- abundance proteins.
  • Fast and easy—<40 min protocol with no centrifugation or preclearing steps.
  • Antibody savings—all binding occurs on the smooth outer surface of the beads, which conserves precious antibodies and supports a cost-efficient solution per sample.
  • Flexible—products for IP, Co-IP, pull-down, and ChIP assays; ideal for manual and automated protocols.
  • Automation of IP protocol—learn how the Thermo Scientific KingFisher platform now supports immunoprecipitation automation.

How to perform Immunoprecipitation using Dynabeads

Step 1: Binding of the antibody to the beads

Binding of the antibody to the beads. Dynabeads magnetic beads are precoated with protein A, protein G, anti-mouse IgG, or anti-rabbit IgG antibodies and due to the rapid kinetics will bind the added antibody in only 10 minutes. Biotinylated antibodies can also be used in combination with streptavidin-coupled Dynabeads. Wash the beads once on the magnet to remove of unbound antibodies.

Step 2: Add sample to the beads

Add sample to the beads. Add the protein-containing sample to the washed beads with bound antibody and incubate for another 10 minutes to bind the target protein of interest. If the protein is of low abundance or low affinity, the incubation can be increased to 1 hour or an overnight incubation; but in general, a 10-minute incubation is sufficient.

Step 3: Wash the protein-bound beads

Wash the protein-bound beads. To ensure high purity of the bound target protein, the beads should be washed 2–4 times with buffer on the magnet to remove all unbound proteins. The efficient isolation process ensures a high signal-to-noise ratio.

Step 4: Elute the protein

Elute the protein. If required, the protein can be eluted off the beads by using either a mild elution procedure or a denaturing elution procedure (for details, see elution description below).

Process and Benefits of IP with Magnetic Dynabeads

Protein elution properties

Protein elution properties
Varying degrees of elution with Dynabeads

The recombinant protein A and/or protein G on the beads contains no albumin binding sites, thus avoiding co-purification of contaminating proteins. Binding of antibodies to the beads in solution is an equilibrium reaction. To capture as many antibodies as possible, the reaction volume should be kept low to maintain high concentrations of beads and antibody. There is no need for pre-treating the sample (even viscous samples), hence no dilution of sample or beads should be done. If the antibody concentration in the sample is suspected to be very low, the amount of beads can be increased.

As an alternative to eluting antibody from the beads, the Dynabeads may be re-suspended in a Na-phosphate buffer. To prevent co-elution, it is possible to cross-link the antibody to protein A/G on the beads prior to immunoprecipitation. This is not necessary for downstream SDS-PAGE followed by autoradiography or western blotting, and optional for silver or coomassie dye staining.

Find the right immunoprecipitation product for your experiment

Choose this if you have an antibody that recognizes your protein

Your choice of antibody binding products depends on the downstream assay.

Antibody binding is the most common method and is used when your target antibody can be bound directly to the beads or indirectly to a pre-coated ligand on the magnetic beads.

Antibody-binding product selection guide

  Protein A, G, A/G Secondary antibodies
(anti-mouse, anti-rabbit)
Surface-activated beads (epoxy)*
Products

Beads only:

Kits:

Beads only:

Kits:

Binding properties Non-covalent antibody binding Non-covalent antibody binding Covalent antibody binding
Antibody co-eluted off the beads Yes Yes No
Type of ligand Different ligands bind different species and antibody subclasses with different specificity Anti-mouse binds mouse IgG1, IgG2a, IgG2b; anti-rabbit binds all rabbit IgGs All antibodies
Mass spec compatible Yes (Protein A/G);
Not validated (Protein A, G)
No Yes
Nonspecific binding Low Low Ultra-low
Antibody coupling time & temperature (RT=room temp.)
  • Protein A: 10 min, RT
  • Protein G: 10–40 min, RT
  • Protein A/G: 60 min, RT
>30 min, 2–8°C 16–24 hr, 37°C

* See more choices in surface-activated Dynabeads for the binding and capture of additional targets.
† Crosslinking can be performed to avoid co-elution of the antibody, but this can decrease the yield of the target antigen.
‡ Learn which antibody-binding proteins are best for your IP antibody.

See ordering information


Choose this if you have a biotinylated antibody (or ligand) that recognizes your protein

Main advantages for using a biotinylated antibody with streptavidin-coated beads for IP include:

  • If you have a sample rich in soluble IgGs
  • If you have a recombinant antibody lacking Fc regions
  • If you already have streptavidin-coated Dynabeads in the lab
  • If you need a bead compatible with mass spectrometry (secondary-coated and epoxy-coated Dynabeads are also compatible with mass spectrometry)

To allow flexibility, four different types of streptavidin-coupled Dynabeads are available. View the Dynabeads streptavidin selection guide to see the different characteristics.

Over the past 25 years, streptavidin-coupled Dynabeads in combination with a biotinylated ligand have been widely used and are cited in thousands of papers for diverse manual and automated applications. The most widely used application for Streptavidin Dynabeads is for purification of nucleic acids and next generation sequencing (NGS) workflows, but they are also used for immunoprecipitation (IP).

See ordering information


Choose this if you have a recombinant (fusion tagged) protein

Popular fusion tags for recombinant protein expression include:

  • His tag—consists of a string of six to nine histidine residues; primarily used for purification via immobilized metal affinity chromatography (IMAC)
  • GST tag—consists of glutathione S-transferase (GST), a complete 211 amino acid protein (26 kDa); primarily used for purification via glutathione agarose resin
  • HA tag—consists of a peptide (YPYDVPDYA) derived from the human influenza hemagglutinin (HA) protein; immobilized anti-HA antibodies are used to immunoprecipitate HA-tagged recombinant proteins
  • c-Myc tag—consists of a peptide (EQKLISEEDL) derived from the human c-myc oncogene (p62 c-myc); immobilized anti-c-Myc antibodies are used to immunoprecipitate c-Myc-tagged recombinant proteins.
  • FLAG tag—consists of a peptide (DYKDDDDK) which is recognized by an immobilized high-affinity rat monoclonal antibody (clone L5); primarily used for the isolation of protein complexes with multiple subunits, because the mild purification process tends not to disrupt these interactions

His and GST are not true epitope tags, because they are not usually purified or detected via specific antibodies. By contrast, HA and c-Myc tags are true epitope tags, because their only means of purification or detection is via specific antibodies. Epitope tags are seldom used for bulk purification but are most often used for immunoprecipitation or co-immunoprecipitation (IP, co-IP). Nevertheless, all four of these tag systems can be used for either purification or pull-down applications.

If you’ve got a biotinylated antibody that recognizes the protein you wish to capture, you can create your own affinity product with one of these products.

See ordering information

Benchmarking data: Dynabeads vs. other IP solutions

Dynabeads continually outperform other methods and magnetic bead solutions for IP. Still using Sepharose or agarose bead slurry. See some of the most common myths about immunoprecipitation.

Dynabeads compared to resin-based solutions

Figure 1. Shorter protocol time and better yields with Dynabeads magnetic beads. Equal amounts of sample material with respect to Ab content and cell lysate volume were used for all IP protocols according to the manufacturer’s protocol. For the Dynabeads-based method, all the antibodies on the bead surface are accessible for optimal, highly reproducible antigen binding.

Dynabeads compared to magnetic solutions

Figure 2. The magnetic bead you choose will affect your results. Dynabeads magnetic beads have a defined surface to carry out the necessary binding, with no inner surface to trap any unwanted proteins. (A) Dynabeads products are the most uniform, monodisperse superparamagnetic beads, manufactured with highly controlled product qualities to enable a high degree of reproducibility. (B–D) Magnetic particles from alternative suppliers have variable shapes and sizes that trap impurities, resulting in lower reproducibility and increased nonspecific binding.

Figure 3. Benchmarking data shows that Dynabeads magnetic products have the best performance when taking into account the combination of highest yield and lowest nonspecific binding. Our magnetic beads also have the fastest protocol, helping improve your lab life by removing unnecessary steps.

Automate immunoprecipitation with KingFisher instruments

Complete protein immunoprecipitation in 40 min with robust and consistent results using Dynabeads on KingFisher instruments.

See how Dynabeads and KingFisher can shorten Immunoprecipitation to 40 min, with only 15 min “hands on” time

KingFisher instruments

Ordering information

Each starter pack has the DynaMag-2 magnet. Choose the starter pack best suited for you.
These bundled combinations save you between 21–24% compared to purchasing separately.

Magnet with Dynabeads only
Magnet with Dynabeads IP Kit

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