Shop All Magnetic Beads Ligand-Coupled

Dynabeads™ M-280 Streptavidin, for OEM and industrial use only

Uniform and superparamagnetic Dynabeads®, 2.8 µm in diameter, with a monolayer of recombinant streptavidin covalently coupled to the surface and further blocked with BSA. The beads allow for efficient capture and downstream handling of your biotinylated ligand or target molecule. The streptavidin monolayer ensures negligible leakage, and the lack of excess adsorbed streptavidin ensures batch consistency and reproducibility of your results.

Streptavidin-coupled Dynabeads® is the gold standard for isolation and handling of biotinylated antibodies, nucleic acids or other biotinylated ligands and targets. The liquid-phase behavior in combination with the superparamagnetic properties of the beads provide rapid reaction kinetics both in the coating process, separation and during washing of the analyte.

The product holds reputable Dynal high standards with respect to reproducibility and automation ability, and drives reliability for your assays.

Benefits and Features:

• Direct and fast capture of any biotinylated ligand or target
• Flexible solid-phase protocols with superior liquid-phase reaction kinetics
• Easy and reproducible handling in manufacturing
• Low charged and neutral beads, optimal for binding of proteins, peptides and antibodies
• Fast and efficient washing procedures
• High batch-to-batch reproducibility, securing consistent results in your application
• Well suited for automated protocols

Applications:
Efficient capture of biotinylated molecules. For direct⁄indirect isolation and downstream handling of nucleic acids, proteins⁄peptides and other target molecules. Due to their uniformity, excellent batch reproducibility and ease of adaptation to automated processes, Dynabeads® have become the solid phase of choice for immunoassay development.

Binding capacity:
The size of the molecule and the biotinylation procedure will affect the binding capacity. The capacity also depends on steric availability and charge interaction between bead and molecule and between molecules. There are two or three biotin binding sites available for each streptavidin molecule on the surface of the bead after immobilization. One mg of Dynabeads® M-280 Streptavidin typically binds 650-900 pmoles free biotin, 200 pmol biotinylated peptides, up to10 µg biotinylated antibody, approx. 10 µg ds-DNA or 200 pmol ss-Oligonucleotides.

Additional Info:
This specific product format is for large volume customers, available on an OEM basis. The product is also available in smaller volumes for end-users (Cat. no 112-05D, 112-06D and 602-10).

Dynabeads™ MyOne™ Streptavidin T1 (Invitrogen™)

Dynabeads MyOne Streptavidin T1 are the gold standard for isolation and handling of biotinylated nucleic acids, antibodies, or other biotinylated ligands and targets. The very high binding affinity of the streptavidin-biotin interaction (Kd=10-15) is used in a vast number of applications. Benefits and features:

• Direct and fast isolation of any biotinylated molecule
• Flexible protocols with gentle and efficient liquid-phase reaction kinetics
• Low-charged and neutral beads, optimal for binding of proteins, peptides, and antibodies
• Their small but uniform size presents a high surface area per mg beads and a correspondingly high capacity for the target molecule
• Low sedimention rate, yet a high iron content, ensuring rapid magnetic separation
• Biomagnetic protocols are easily adapted to automated platforms
• High batch-to-batch reproducibility, securing consistent results in your application

About Dynabeads MyOne Streptavidin T1
These uniform and superparamagnetic beads are 1 µm in diameter, with a monolayer, not a multilayer, of recombinant streptavidin covalently coupled to the surface and further blocked with BSA. The monolayer of streptavidin leaves the vast majority of the biotin binding sites sterically available for binding, not only of free biotin, but also for binding of biotinylated ligands/targets. The beads show rapid liquid-phase reaction kinetics. Their specific and defined surface allow for efficient capture, separation, and downstream handling. The streptavidin monolayer ensures negligible leakage, and the lack of excess adsorbed streptavidin ensures batch consistency and reproducibility of your results. The 1 µm Dynabeads MyOne have a large surface area, high capacity, efficient magnetic pull, and a slow sedimentation rate during incubation. Tailor-made for use in automated protocols where high throughput is crucial.

Applications
Over the past 15 years, streptavidin-coupled Dynabeads have been used and cited for a very wide variety of applications. Key applications include preparing single-stranded DNA templates, isolation of RNA and DNA binding proteins, immobilization of large DNA fragments, purifying sequencing products, and the specific capture of nucleic acids. Easily adapted to automated processes. Dynabeads are used on more than 25,000 routine IVD instruments worldwide.

Binding capacity
The size of the molecule and the biotinylation procedure will affect the binding capacity. The capacity also depends on steric availability and charge interaction between bead and molecule and between molecules. There are two or three biotin binding sites available for each streptavidin molecule on the surface of the bead after immobilization. One mg of Dynabeads MyOne Streptavidin T1 typically binds:

• 950–1500 pmoles free biotin
• ~20 µg biotinylated IgG
• ~400 pmol biotinylated peptides
• ~20 µg ds-DNA
• ~400 pmol ss-oligonucleotides

Dynabeads™ Protein G and Magnet Starter Pack (Invitrogen™)

The Dynabeads Protein G and Magnet Starter Pack combines Dynabeads Protein G magnetic beads for immunoprecipitation with the magnet preferred for use with 1.5 mL microcentrifuge tubes for purchasing convenience. The beads are sufficient for 100 reactions, and the magnet holds 16 x 1.5 mL microcentrifuge tubes. Dynabeads Protein G beads and the DynaMag-2 magnet are also available separately.

The product contains:
5 mL Dynabeads Protein G beads
1 x DynaMag-2 magnet

Dynabeads™ M-280 Streptavidin (Invitrogen™)

Dynabeads® M-280 Streptavidin are the gold standard for the isolation and handling of biotinylated nucleic acids, antibodies, or other biotinylated ligands and targets. The strong binding affinity of the streptavidin-biotin interaction (Kd=10-15) is used in a vast number of applications.

Benefits and features include:

• Direct and fast isolation of any biotinylated molecule
• Low-charged and neutral beads, optimal for binding of DNA fragments, proteins, peptides, and antibodies
• Flexible protocols with gentle and efficient liquid-phase reaction kinetics
• Biomagnetic protocols are easily adapted to automated platforms
• High batch-to-batch reproducibility, securing consistent results in your application

About Dynabeads® M-280 Streptavidin
These uniform and superparamagnetic beads are 2.8 µm in diameter, with a monolayer, not a multilayer, of recombinant streptavidin covalently coupled to the surface and further blocked with BSA. The monolayer of streptavidin leaves the vast majority of the biotin binding sites sterically available for binding, not only of free biotin, but also for binding of biotinylated ligands/targets. They show rapid liquid-phase reaction kinetics. Their specific and defined surface allow for efficient capture, separation, and downstream handling. The streptavidin monolayer ensures negligible leakage, and the lack of excess adsorbed streptavidin ensures batch consistency and reproducibility of your results.

Applications
Over the past 15 years, Dynabeads® M-280 Streptavidin have been used and cited for a very wide variety of applications. Key applications include preparing 2–5 kb samples for mate pair library sequencing using Illumina platforms preparing single-stranded DNA templates, isolation of RNA and DNA binding proteins, immobilization of large DNA fragments, purifying sequencing products, and the specific capture of nucleic acids. Dynabeads® are used on more than 25,000 routine IVD instruments worldwide. The product holds high standards with respect to reproducibility (both within and between batches), and automation ability, and drives reliability for your results.

Binding capacity
The size of the molecule and the biotinylation procedure will affect the binding capacity. The capacity also depends on steric availability and charge interaction between bead and molecule and between molecules. There are two or three biotin binding sites available for each streptavidin molecule on the surface of the bead after immobilization. One mg of Dynabeads® M-280 Streptavidin typically binds:

• 650–900 pmoles free biotin
• ~200 pmol biotinylated peptides
• ~10 µg biotinylated IgG
• ~10 µg ds-DNA
• ~200 pmol ss-oligonucleotides

Pierce™ Anti-HA Magnetic Beads (Thermo Scientific™)

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.

Features of Anti-HA Magnetic Beads:

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)

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.

Specifications:
Product Details:
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, 50 mM NaOH, or SDS-PAGE sample buffer. If gentler elution conditions are desired, 2 mg/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.

Dynabeads™ Protein G for Immunoprecipitation (Invitrogen™)

Dynabeads™ Protein G are uniform, 2.8 µm superparamagnetic beads with recombinant Protein G (~17 kDa) covalently coupled to the surface. Dynabeads Protein G provide a superior alternative to Sepharose™ or agarose slurry for immunoprecipitation (IP), and both manual and automated protocols are available.

• IP in less than 40 minutes
• High target protein yield with low antibody consumption
• Very low non-specific binding with high signal-to-noise ratio
• No columns, centrifugations, or time-consuming pre-clearing required
• High reproducibility and high throughput compatible with KingFisher™ instruments

Manual Dynabeads separation is fast and easy to perform
The manual protocol is simple and can be performed in under 40 minutes. First, the antibody for the target protein is incubated with the Dynabeads Protein G in a tube for 10 minutes. Excess antibody is washed away by placing the tube in a DynaMag™ magnet and removing the supernatant. The antibody-coated beads can then be used for a variety of downstream applications including IP, Co-IP, chromatin IP (ChIP), RNA IP (RIP), small-scale IgG purification, and protein purification. Bound material is easily collected using a DynaMag magnet due to the unique magnetic properties of the Dynabeads. The recombinant protein G on the beads contains no albumin binding sites, thus albumin is not co-purified during the procedure. The IP is fast and gives high yield, high reproducibility, and very little non-specific binding, thus pre-clearing is not required.

Automated Dynabeads separation helps increase throughput and reduces hands-on time
If you are working with several samples in parallel, the number of washing steps and the hands-on time increases proportionally with the number of samples. Pipetting and other manual handling tend to be less consistent than automation when working with many samples at a time. To better handle a medium- to high-throughput number of samples, reduce hands-on time, and secure high reproducibility, we have developed IP protocols for the KingFisher Flex and KingFisher Duo Prime instruments. The automated protocols replicate the manual protocols, obtaining equally high target protein yield and the same low non-specific binding and high reproducibility. It doesn’t matter if you are working with 10 or 96 samples, the IP protocol is less than 40 minutes regardless. Just load the reagents on the plates, push the “Start” button and by the time you have prepared for downstream analysis, the IP is done. Some optimization (e.g., incubation times) might be necessary depending on your antibody and the abundance and/or specificity of your target protein.

• Use the KingFisher Duo instrument for low to medium throughput (1-12 samples/run)
• Use the KingFisher Flex instrument for high throughput (12-96 samples/run)
See automated protocols
Watch a video about the KingFisher Flex instrument

Gentle separation causes minimal physical stress to proteins
The magnetic separation technology utilized by Dynabeads Protein G is rapid and gentle, causing minimal physical stress to your target proteins. This permits the isolation and concentration of labile composites that might otherwise dissociate or be damaged by proteases during long incubation times. Native protein conformation and large protein complexes are preserved.

Binding strength and capacity
Dynabeads Protein G allow for isolation of most mammalian immunoglobulins (Ig). The amount of Ig captured depends on the concentration of Ig in the starting sample and on the type and source of the Ig. 100 µL of Dynabeads Protein G will isolate approximately 25–30 µg human IgG from a sample containing 20–200 µg IgG/mL. Predominant Fc-binding allows optimal Ig orientation. The antibodies bind to the outer smooth surface of the beads, thus are not trapped in large pores as with Sepharose/agarose-based beads. All antibodies are available for protein binding, so low amounts of antibody are required while still obtaining the same high yield of target protein. The smooth bead surface is also responsible for the low non-specific binding that Dynabeads are known for.

Learn more about Dynabeads
• Dynabeads Protein A are also available as a "ready-to-go" kit with buffers included
See immunoprecipitation selection guides, data, and references
See magnets for Dynabeads separations
Find Dynabeads products for other applications

OEM purchase
To purchase Dynabeads Protein A and Protein G on an OEM basis, contact our Out-Licensing and OEM Sales department.

*Sepharose is a trademark of GE Healthcare Bio-Sciences AB.

MagnaBind™ Protein G Beads (Thermo Scientific™)

Thermo Scientific™ MagnaBind™ Beads provide a convenient method for magnetic separation of antibodies, antigens, lectins, enzymes, nucleic acids and cells using affinity binding. To remove the MagnaBind Beads from the suspension, an external magnetic field is used.

Features of MagnaBind Protein G Beads:

Composition: Silanized iron oxide
Magnetization: 25-35EMU/g
Type of Magnetization: Superparamagnetic (no magnetic memory)
Surface Area: >100m2/g
Bead Size: 1-4µm diameter
Settling Rate: 4% in 30 minutes
Effective Density: 2.5g/mL
Number of Beads: 1 × 108 beads/mg
pH Stability: Aqueous solution, above pH 4.0
Concentration: 5 mg/mL

MagnaBind Protein G Beads are typically used to isolate antibodies from serum and cell culture supernatant and to separate cells of interest from a cell mixture. For antibody purification, the beads are incubated with the antibody solution and then magnetically separated from the supernatant. The antigens and antibodies are then dissociated from the beads using an elution buffer. For cell separation, the monoclonal or polyclonal antibody to the cell surface antigen is incubated with Protein G magnetic beads, magnetically separated, and then incubated with the cell suspension.

Applications:
• Cell sorting using either positive or negative selection
• Protein purification or immunoassays using either direct or indirect methods

Dynabeads™ M-280 Sheep Anti-Rabbit IgG (Invitrogen™)

Dynabeads™ M-280 Sheep Anti-Rabbit IgG are 2.8 µm superparamagnetic beads with affinity-purified polyclonal sheep anti-rabbit IgG covalently bound to the bead surface. These beads are designed to serve as a solid support for simple and efficient binding of rabbit IgGs of all subclasses and their target proteins. The uniform, monodisperse, and non-porous Dynabeads make them an ideal choice for applications such as binding Ig, protein purification, sandwich immunoassays, immunoprecipitation (IP), Co-IP, and isolation of cells and microorganisms.

• Isolate protein in less than 40 minutes
• Binds all rabbit IgG subclasses
• Very low non-specific binding with high signal-to-noise ratio
• No columns, centrifugations, or time-consuming pre-clearing required
• High reproducibility and high throughput compatible with KingFisher™ instruments

Manual Dynabeads separation is fast and easy to perform
The primary antibody that recognizes the target molecule may be added to the sample (indirect technique) or pre-coated onto the Dynabeads M-280 Sheep Anti-Rabbit IgG (direct technique). Regardless of which technique is used, when the Dynabeads M-280 Sheep Anti-Rabbit IgG are mixed with the sample, the beads bind to the target. Placing the sample on a DynaMag™ magnet separates the bead-bound target from the rest of the sample. The supernatant is then removed by aspiration. The target molecule can be eluted off the beads with conventional elution methods and used in such applications as Western blot or mass spec analysis, or used in further downstream applications while still attached to the beads.

Automated Dynabeads separation helps increase throughput and reduces hands-on time
If you are working with several samples in parallel, the number of washing steps and the hands-on time increases proportionally with the number of samples. Pipetting and other manual handling tend to be less consistent than automation when working with many samples at a time. To better handle a medium- to high-throughput number of samples, reduce hands-on time, and secure high reproducibility, we have developed IP protocols for the KingFisher Flex and KingFisher Duo Prime instruments. The automated protocols replicate the manual protocols, obtaining equally high target protein yield and the same low non-specific binding and high reproducibility. It doesn’t matter if you are working with 10 or 96 samples, the IP protocol is less than 40 minutes regardless. Just load the reagents on the plates, push the “Start” button and by the time you have prepared for downstream analysis, the IP is done. Some optimization (e.g., incubation times) might be necessary depending on your antibody and the abundance and/or specificity of your target protein.

• Use the KingFisher Duo instrument for low to medium throughput (1-12 samples/run)
• Use the KingFisher Flex instrument for high throughput (12-96 samples/run)
See automated protocols
Watch a video about the KingFisher Flex instrument

Learn more about Dynabeads
See immunoprecipitation selection guides, data, and references
See magnets for Dynabeads separations
Find Dynabeads products for other applications

OEM purchase
To purchase Dynabeads Sheep Anti-Rabbit IgG on an OEM basis, contact our Out-Licensing and OEM Sales department.

HisPur™ Ni-NTA Magnetic Beads (Thermo Scientific™)

Thermo Scientific HisPur Ni-NTA Magnetic Beads are high-capacity nickel-IMAC beads for affinity purification of His-tagged fusion proteins in manual or automated formats.

Features of HisPur Ni-NTA Magnetic Beads:

High capacity—equivalent or higher binding capacity than Ni-NTA magnetic beads from other suppliers
Low nonspecific binding—the bead surface is pre-blocked and the protocol provides optimized buffers for purification
Fast—protocol is completed in 1 hour
Scalable—process microliter to milliliter sample volumes
Versatile—purify proteins using native or denaturing conditions
Reagent compatible—can be used with common cell lysis reagents and a variety of buffer additives
Multiple formats—protein coupling to the beads and downstream applications can be performed both manually and on an automated platform (e.g., Thermo Scientific KingFisher Instruments)

The blocked magnetic bead surface is derivatized with the nitrilotriacetic acid (NTA) chelation moiety and loaded with divalent nickel ions (Ni2+). The immobilized metal affinity chromatography (IMAC) beads provide high binding capacity with very low background. The HisPur Ni-NTA Magnetic Beads can be used both manually with a magnetic stand as well as with automated platforms such as the Thermo Scientific KingFisher Instruments for high-throughput needs.

HisPur Ni-NTA Magnetic Beads are used for small scale affinity purification as well as high-throughput screening of recombinant His-tagged proteins. The polyhistidine tag is the most popular affinity tag and typically consists of six consecutive histidine residues (6xHis). These tagged proteins are overexpressed in a number of different systems, most commonly in bacteria, and purified from cell lysates such as those prepared using B-PER Bacterial Protein Extraction Reagents. Purification of His-tagged proteins is achieved using a NTA chelate charged with nickel that coordinates with the histidine side chains. The NTA chelate contains four metal-binding sites which allow for low metal ion leaching and high binding capacity. The protocol for the HisPur Ni-NTA Magnetic Beads has been optimized to allow for high purity of the isolated His-tagged protein. Performance is equivalent to or better than Ni-NTA magnetic beads from other suppliers.

MagnaBind™ Goat Anti-Rabbit IgG (Thermo Scientific™)

Thermo Scientific™ MagnaBind™ Beads provide a convenient method for magnetic separation of antibodies, antigens, lectins, enzymes, nucleic acids and cells using affinity binding. To remove the MagnaBind Beads from the suspension, an external magnetic field is used.

MagnaBind Goat Anti-Rabbit IgG Beads cab be used to conveniently separate cells of interest from a cell mixture. The monoclonal or polyclonal antibody to the cell surface antigen is pre-incubated with the appropriate magnetic bead that is then incubated with the cell suspension.

Features of MagnaBind Goat Anti-Rabbit IgG Beads:

Composition: Silanized iron oxide
Magnetization: 25-35EMU/g
Type of Magnetization: Superparamagnetic (no magnetic memory)
Surface Area: >100m2/g
Bead Size: 1-4µm diameter
Settling Rate: 4% in 30 minutes
Effective Density: 2.5g/mL
Number of Beads: 1 × 108 beads/mg
pH Stability: Aqueous solution, above pH 4.0
Concentration: ~1 mg/mL

Applications:
• Cell sorting using either positive or negative selection
• Protein purification or immunoassays using either direct or indirect methods

Dynabeads™ Protein A and Magnet Starter Pack (Invitrogen™)

The Dynabeads Protein A and Magnet Starter Pack combines Dynabeads Protein A magnetic beads for immunoprecipitation with the magnet preferred for use with 1.5 mL microcentrifuge tubes for purchasing convenience. The beads are sufficient for 100 reactions, and the magnet holds 16 x 1.5 mL microcentrifuge tubes. Dynabeads Protein A beads and the DynaMag-2 magnet are also available separately.

The product contains:
5 mL Dynabeads Protein A beads
1 x DynaMag-2 magnet

Pierce™ Streptavidin Magnetic Beads (Thermo Scientific™)

Thermo Scientific Pierce Streptavidin Magnetic Beads accelerate throughput for automated magnetic purification of biotinylated molecules.

Features of Streptavidin Magnetic Beads:

High-performance beads—non-aggregating, pre-blocked, iron oxide, superparamagnetic microparticles provide exceptional uniformity for automated HTS and manual applications alike
Stable immobilization chemistry—streptavidin is immobilized using leach-resistant chemistry
High capacity—superior quality beads with high binding capacity provide rapid and efficient biomolecule purification from complex samples
Low non-specific binding—stable, pre-blocked beads provide clean purification products (e.g., antigen eluted in IP with biotinylated antibody) that are compatible with mass spectrometry analysis and other downstream analyses
Superior performance—nearly three times higher binding capacity than typical beads from other suppliers, allowing the use of smaller amounts per experiment

Applications:
• Immunoprecipitate antigens (using biotinylated antibodies) from a wide variety of sources
• Co-immunoprecipitate interaction complexes using biotinylated antibodies
• Capture protein-protein interactions in pull-down assays using biotinylated "bait" proteins
• Isolate biotin-labeled DNA-protein complexes from cell or tissue extracts
• Capture single-stranded biotinylated DNA oligos
• Isolate biotinylated PCR products

These streptavidin magnetic beads are validated and optimized for use with high-throughput magnetic platforms, such as the Thermo Scientific KingFisher 96 and KingFisher Flex Instruments, but the beads also enable premium performance for simple benchtop applications using an appropriate magnetic stand. The iron oxide, super-paramagnetic particles offer superior performance (high capacity and low nonspecific binding) compared with other commercial magnetic beads.

Pierce Streptavidin Magnetic Beads use a recombinant form of streptavidin with a mass of 53kDa and a near-neutral isoelectric point (pI). The protein is a tetramer having four biotin-binding sites. Unlike avidin, streptavidin has no carbohydrate groups, resulting in low nonspecific binding. The high-affinity interaction between streptavidin and biotin cannot be dissociated efficiently except with very harsh conditions, such as boiling in sample loading buffer for SDS-PAGE or 8M guanidine·HCl, pH 1.5. Consequently, it is often possible to elute binding partners in an interaction complex without also eluting the biotinylated component.

Exosome Immunoprecipitation Reagent (Protein G) (Invitrogen™)

Exosome Immunoprecipitation Reagent (Protein G) enables fast and gentle magnetic isolation of exosomal proteins, causing minimal physical stress to the target protein and allowing comparison of multiple samples on the same gel.

• Maintain intact exosome protein complexes
• Reduce background significantly—low non-specific binding
• Fast protocol time—only 30 minutes
• Maximal comparison—ultra-sensitivity allows many samples on the same gel

Dynabeads® coupled to Protein G are widely used for immunoprecipitation (IP), chromatin immunoprecipitation (ChIP), and protein isolation. The magnetic properties of Dynabeads® makes them a superior alternative to sepharose or agarose slurry for immunoprecipitation, since magnetic separation technology is faster and gentler than other methods, causing minimal physical stress to the target proteins. This permits the isolation and concentration of labile composites that might otherwise dissociate or be damaged by proteases during long incubation times. Native protein conformation and intact, large protein complexes will be preserved.

Exosome Immunoprecipitation (Protein G) makes use of Dynabeads® Protein G for antibody binding and subsequent immunoprecipitation of exosomal proteins. Antibody is added to the Dynabeads® suspension where binding occurs via the Fc-region of the antibody. The mixture is then placed near a magnet, causing the beads migrate to the side of the tube, allowing easy removal of the supernatant. The bead-bound antibody can now be used for immunoprecipitation of exosomal proteins. Immunoprecipitation allows a 10 to 50 times concentration of exosomal proteins prior to protein analysis, such as Western blotting.

The amount of Ig captured by Exosome Immunoprecipitation (Protein G) is dependent on the concentration of Ig in the starting sample. The binding capacity is approximately 240 µg human Ig per mL beads.

For isolation of Ig via protein A, we recommend the Exosome Immunoprecipitation (Protein A).

For Research Use Only. Not for human or animal therapeutic or diagnostic use.

Pierce™ Protein A Magnetic Beads (Thermo Scientific™)

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

Features of Protein A Magnetic Beads:

High efficiency – equivalent or higher yield of IP target antigens than magnetic beads from other suppliers
Low non-specific binding – stable, pre-blocked beads provide highly purified product (e.g., antigen eluted in IP with antibody is devoid of contaminating protein from complex cell lysate)
Consistent – magnetic beads eliminate resin loss and provide for more efficient separation than traditional IP methods that use only centrifugation
Versatile – beads are compatible with manual and automated workflows (e.g., Thermo Scientific KingFisher Instruments)

Recombinant protein A, covalently attached to a blocked magnetic bead surface, can bind to antibodies from many different species, enabling purification of antibodies from crude extracts. Immunoprecipitation assays performed with protein A coated beads result in high yield of target antigen with very low background. Using our crosslinker chemistry, you can immobilize an antibody onto the magnetic particle and prevent IgG contamination in IP or Co-IP experiments. The Pierce Protein A 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.

Applications:
• IP and Co-IP experiments
• Antibody purification

Thermo Scientific Pierce Protein A Magnetic Beads are used for purifying antibody from serum, cell culture supernatant or ascites, as well as for IP/Co-IP of antigens from cell or tissue extracts. Protein A can bind to antibodies from many different species, including mouse, human, rabbit, pig, dog, and cat. The protocol for the Pierce Protein A beads has been optimized to allow for high recovery and high purity of the isolated antibody or antigen. Antibody or antigen/antibody complex (IP) is first captured on the magnetic beads. The beads are washed and then the target is eluted with low pH elution buffer. IP performance is equivalent to or better than Protein A magnetic beads from other suppliers.

Dynabeads™ Protein G Immunoprecipitation Kit (Invitrogen™)

The Dynabeads™ Protein G Immunoprecipitation Kit is a faster and easier solution for immunoprecipitation (IP) than using Sepharose™ resin or agarose resin, and includes all reagents and buffers required to perform IP using your own antibody. The kit is optimized for standard IP, but can also be used for Co-IP, chromatin IP (ChIP), or RNA IP (RIP). The kit includes the widely used Dynabeads Protein G with recombinant Protein G (~17 kDa) covalently coupled to the magnetic bead surface. Both manual and automated protocols are available.

• IP in less than 40 minutes
• High target protein yield with low antibody consumption
• Very low non-specific binding with high signal-to-noise ratio
• No columns, centrifugations, or time-consuming pre-clearing required
• High reproducibility with Dynabeads Protein G and all buffers supplied in the kit
• High throughput compatible with KingFisher™ instruments

Manual Dynabeads separation is fast and easy to perform
The manual protocol is simple and can be performed in under 40 minutes. First, the antibody for the target protein is incubated with the Dynabeads Protein G in a tube for 10 minutes. Excess antibody is washed away by placing the tube in a DynaMag™ magnet and removing the supernatant. The antibody-coated beads can then be used for a variety of downstream applications including IP, Co-IP, chromatin IP (ChIP), RNA IP (RIP), small-scale IgG purification, and protein purification. Bound material is easily collected using a DynaMag magnet due to the unique magnetic properties of the Dynabeads. The recombinant protein G on the beads contains no albumin binding sites, thus albumin is not co-purified during the procedure. The IP is fast and gives high yield, high reproducibility, and very little non-specific binding, thus pre-clearing is not required.

Automated Dynabeads separation helps increase throughput and reduces hands-on time
If you are working with several samples in parallel, the number of washing steps and the hands-on time increases proportionally with the number of samples. Pipetting and other manual handling tend to be less consistent than automation when working with many samples at a time. To better handle a medium- to high-throughput number of samples, reduce hands-on time, and secure high reproducibility, we have developed IP protocols for the KingFisher Flex and KingFisher Duo Prime instruments. The automated protocols replicate the manual protocols, obtaining equally high target protein yield and the same low non-specific binding and high reproducibility. It doesn’t matter if you are working with 10 or 96 samples, the IP protocol is less than 40 minutes regardless. Just load the reagents on the plates, push the “Start” button and by the time you have prepared for downstream analysis, the IP is done. Some optimization (e.g., incubation times) might be necessary depending on your antibody and the abundance and/or specificity of your target protein.

• Use the KingFisher Duo instrument for low to medium throughput (1-12 samples/run)
• Use the KingFisher Flex instrument for high throughput (12-96 samples/run)
See automated protocols
Watch a video about the KingFisher Flex instrument

Gentle separation causes minimal physical stress to proteins
The magnetic separation technology utilized by Dynabeads Protein G is rapid and gentle, causing minimal physical stress to your target proteins. This permits the isolation and concentration of labile composites that might otherwise dissociate or be damaged by proteases during long incubation times. Native protein conformation and large protein complexes are preserved.

Binding strength and capacity
Dynabeads Protein G allow for isolation of most mammalian immunoglobulins (Ig). The amount of Ig captured depends on the concentration of Ig in the starting sample and on the type and source of the Ig. 100 µL of Dynabeads Protein G will isolate approximately 25–30 µg human IgG from a sample containing 20–200 µg IgG/mL. Predominant Fc-binding allows optimal Ig orientation. The antibodies bind to the outer smooth surface of the beads, thus are not trapped in large pores as with Sepharose/agarose-based beads. All antibodies are available for protein binding, so low amounts of antibody are required while still obtaining the same high yield of target protein. The smooth bead surface is also responsible for the low non-specific binding that Dynabeads are known for.

Learn more about Dynabeads
• Dynabeads Protein G are also available as a stand-alone reagent without included buffers
See immunoprecipitation selection guides, data, and references
See magnets for Dynabeads separations
Find Dynabeads products for other applications

OEM purchase
To purchase Dynabeads Protein A and Protein G on an OEM basis, contact our Out-Licensing and OEM Sales department.

*Sepharose is a trademark of GE Healthcare Bio-Sciences AB.