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

The Dynabeads Protein A IP Kit and Magnet Starter Pack combines Dynabeads Protein A magnetic beads for immunoprecipitation (including the required buffers) and the magnet preferred for use with 1.5 mL microcentrifuge tubes for purchasing convenience. The beads and buffers are sufficient for 40 reactions, and the magnet holds 16 x 1.5 mL microcentrifuge tubes. The Dynabeads Immunoprecipitation Kit - Protein A with buffers, stand-alone Dynabeads Protein A beads, and the DynaMag-2 magnet are also available separately.

The product contains:
1 x Dynabeads Immunoprecipitation Kit - Protein A*
1 x DynaMag-2 Magnet

*The kit contains 2 mL Dynabeads Protein A beads, 16 mL Antibody Binding & Washing Buffer, 28 mL Washing Buffer, and 1 mL Elution Buffer.

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™ 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.

Exosome Immunoprecipitation Reagent (Protein A) (Invitrogen™)

Exosome Immunoprecipitation Reagent (Protein A) 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 A 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 A) makes use of Dynabeads® Protein A 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 A) is dependent on the concentration of Ig in the starting sample. The binding capacity is approximately 240 ug human Ig per mL beads.

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

Dynabeads™ Protein A Immunoprecipitation Kit (Invitrogen™)

The Dynabeads™ Protein A 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 A with recombinant Protein A (~45 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 A 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 A 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 A 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 A 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 A 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 A 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 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.

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.

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.

MagniSort™ Streptavidin Negative Selection Beads (Invitrogen™)

MagniSort™ Streptavidin Negative Selection Beads are designed for the magnetic separation of cells by negative selection. Undesired cells are bound by a cocktail of biotinylated antibodies and then streptavidin coated magnetic beads. When placed in a magnetic field, the undesired cells are held in place and the desired cells can be separated by decanting. The clone and concentration of biotinylated antibody to be used to label the undesired cells, as well as the concentration of MagniSort™ Streptavidin Negative Selection Beads must be determined empirically. Please refer to the protocol for details on how to optimize both the antibody or antibody cocktail and MagniSort™ Streptavidin Negative Selection Beads.

For depletion of cells using a single antibody, we recoomend the MagniSort™ Streptavidin Positive Selection Beads (cat. MSPB-6003) and protocol.

Reported Application
Magnetic Cell Separation

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

The Dynabeads Protein A/Protein G and Magnet Starter Pack combines Dynabeads Protein A and Dynabeads Protein G magnetic beads for immunoprecipitation with the magnet preferred for use with 1.5 mL microcentrifuge tubes for purchasing convenience. This pack is ideal if you don't know whether Protein A or Protein G is best for your application or if you prefer to use a combination of both. The beads are sufficient for 40 reactions, and the magnet holds 16 x 1.5 mL microcentrifuge tubes. Dynabeads Protein A beads, Dynabeads Protein G beads, and the DynaMag-2 magnet are also available separately.

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