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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.

Dynabeads™ GC-Combo (Applied Biosystems™)

For the selective enrichment of Giardia cysts and Cryptosporidium oocysts directly from water sample concentrates.

Performance tested: Validated IMS in the US Environment Protection Agency's Method 1623.

Benefits and features

• Significant improved recovery of Cryptosporidium & Giardia
• Simple procedure
• Reduced background debris aids detection by microscopy
• One clean 9mm slide well per sample
• Saves hours of hands-on processing time

This Product Contains: Dynabeads® coated with either anti-Cryptosporidium or anti-Giardia antibodies and SL buffer

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 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
• 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

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 A for Immunoprecipitation (Invitrogen™)

Dynabeads™ Protein A are uniform, 2.8 µm superparamagnetic beads with recombinant Protein A (~45 kDa) covalently coupled to the surface. Dynabeads Protein A provide a superior alternative to Sepharose™ resin or agarose resin 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 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 "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.

CTS™ (Cell Therapy Systems) Dynabeads™ CD3/CD28 (Gibco™)

CTS™ Dynabeads™ CD3/CD28 are intended for ex vivo isolation, activation, and expansion of human T-cells in translational research. The technology—previously known as Xcyte™ Dynabeads™ or Dynabeads™ ClinExVivo™ CD3/CD28—has been used in a number of clinical studies. By combining anti-CD3 and anti-CD28 antibodies on Dynabeads, the beads provide both the primary and co-stimulatory signals that are required for activation and expansion of T cells. CTS Dynabeads CD3/CD28 are manufactured and controlled according to ISO 9001:2000 and ISO 13485:2012 requirements. A Device Master File is held with the FDA for cross-referencing in IND applications, and a CoA is available on request. In the US, CTS Dynabeads CD3/CD28 are available for clinical use only under an approved Investigational New Drug (IND) application. Dynabeads™ Human T-Expander CD3/CD28 (Cat. No. 11141D) is the research grade version of CTS Dynabeads CD3/CD28, containing the same amount and ratio of antibodies from the same clones as CTS Dynabeads CD3/CD28, and intended for small scale pre-clinical research.

Applications:
The CTS Dynabeads CD3/CD28 technology has been successfully used in ex vivo isolation, activation, and expansion of T-cells in a number of studies, including studies on HIV infection and cancer. Other potential applications for this novel technology include research into treatment of autoimmune and infectious diseases and prevention of complications associated with chemotherapy and allogeneic stem cell transplantation.

Dynabeads™ MyOne™ Streptavidin C1 (Invitrogen™)

Dynabeads® MyOne™ Streptavidin C1 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
• Well suited for nucleic acid applications with extreme demands
• Very low nonspecific binding of nucleotides and nucleic acids
• Low nonspecific binding of small and negatively charged proteins
• 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
• Production follows a validated process in compliance with cGMP for medical devices

About Dynabeads® MyOne™ Streptavidin C1
These uniform and superparamagnetic beads are 1 µm in diameter, with a monolayer, not a multilayer, of recombinant streptavidin covalently coupled to the surface. This 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 are hydrophilic, negatively charged, and 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. The product holds high standards with respect to reproducibility (both within and between batches), and automation ability, and drives reliability for your results.

Applications
Over the past 15 years, Streptavidin-coupled Dynabeads® have been used and cited for a very wide variety of applications. Examples include direct/indirect isolation and downstream handling of nucleic acids, proteins/peptides and other target molecules. Ideal for sequence specific DNA/RNA capture in nucleic acid based diagnostics, specifically with samples with a high chaotropic salt concentration, immunoassays involving small biotinylated antigens and applications that are not compatible with BSA (these beads are not blocked with BSA). Easily adapted to automated processes. Did you know that Dynabeads® are employed 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 C1 typically binds:

• >2,500 pmoles free biotin
• ~20 µg biotinylated IgG
• ~400 pmol biotinylated peptides
• ~20 µg ds-DNA
• ~500 pmol ss-oligonucleotides

Dynabeads™ MyOne™ Silane

An excellent tool for highly predictable and consistent extraction and isolation of nucleic acids from biological samples, following a simple magnetic separation protocol. Please note that this is a stand-alone product containing only Dynabeads® in a storage buffer. Our products Dynabeads® SILANE genomic DNA (Cat.no. 370-12D) or Dynabeads® SILANE viral NA (Cat.no. 370-11D) include the appropriate buffers and protocols you need to extract genomic DNA or viral nucleic acids respectively.

Magnetic particles from alternative suppliers often have a random size range distribution, surface area and binding capacity. This could compromise the reproducibility of your results. With Dynabeads® MyOne™ SILANE you are ensured a higher sensitivity, capacity and performance.

The small 1 µm Dynabeads® MyOne™ SILANE have an optimized silica-like surface chemistry and a high specific surface area, providing efficient kinetics and a high sensitivity in nucleic acid capture. Their uniform size and surface area also ensure reproducible results. The product holds reputable Dynal® high standards with respect to within- and between-lot reproducibility and automation ability.

Advantages:

• Highly predictable binding per mg of beads
• High sensitivity, allowing for a low detection limit
• For isolation of viral DNA⁄RNA, total RNA, genomic DNA, etc.
• Isolates both DNA and RNA
• Quicker than spin columns
• More cost-effective, yet performs to the same level as spin columns
• Automation-friendly (slow sedimentation rate + high magnetic mobility)
• Adjusted to customer requirements on a custom OEM basis

Applications:
• Isolation of total nucleic acids (DNA⁄RNA)from typical biological samples (e.g. serum⁄plasma, whole blood, etc.)
• Well suited for automated assays.
• Dedicated kits have been developed for isolation of viral nucleic acids and genomic DNA (available separately).

Additional Info:
Validation and customization (bead, buffer, protocol or format) can be made available on a custom OEM basis. For further information and price quotes for larger volumes of the silica-like Dynabeads® MyOne™ SILANE, please contact us at: ivd@invitrogen.com.

Dynabeads™ Oligo(dT)25 (Invitrogen™)

Dynabeads® Oligo(dT)25 mRNA isolation beads specifically target and capture mRNA molecules from virtually any crude sample and eliminate the need to purify total RNA when the desired information-bearing nucleic acid is mRNA. Since mRNA comprises only about 1–5% ot total cellular RNA, the isolation of total RNA is not the most efficient way to isolate mRNA. Other technologies designed to purify total RNA yield ~80% ribosomal RNA and force mRNA to compete with ribosomal RNA, transfer RNA, micro RNA, small nucleolar RNA, and small cytoplasmic RNA for membrane binding. Advantages of Dynabeads® Oligo(dT)25 beads:

• Fast and gentle procedure yields pure intact mRNA
• Extremely pure mRNA isolation, best choice upstream of cDNA synthesis
• Exquisitely sensitive mRNA isolation enables cDNA synthesis and cDNA library construction from ultra-small starting samples (enables cDNA library construction from a single cell)

How the beads work
The oligo(dT)25-coated Dynabeads® specifically target and capture the mRNA transcriptome from an extremely wide variety of crude starting samples. Ribosomal RNA, DNA, proteins, and small RNA molecules (such as transfer RNA, micro RNA, and small nucleolar RNA) do not bind to the beads and are discarded. Only polyadenylated RNA species (mRNA) are captured. Isolated mRNA is pure, eliminating the need for ribosomal RNA subtraction or a post-extraction DNase treatment. This column-free system ensures the highest transcriptome recovery:

• Physical mRNA capture on mobile magnetic beads
• Rapid and gentle magnetic handling procedures
• No mRNA lost during high g-force spins
• No mRNA trapped in column membranes during elution

Applications
mRNA is suitable for all downstream molecular applications, including gene cloning, cDNA synthesis, cDNA library construction, RT-PCR, quantitative RT-PCR, RPA (Ribonuclease Protection Assay), subtractive hybridization, primer extension, SAGE, RACE, and others. The Dynabeads® Oligo(dT)25 mRNA isolation beads are the ideal mRNA purification method prior to cDNA library construction. Use of these beads ensures the highest recovery and enrichment of the transcriptome. These beads capture more of the transcriptome than is possible with methods that integrate a total RNA isolation step upstream of mRNA isolation.

Verastile elution options
Elution can be performed in any volume down to 5 µL. mRNA elution is optional because enzymatic reactions in downstream procedures are not inhibited by presence of Dynabeads®. Additionally, one can perform cDNA synthesis directly on the beads to create a reusable solid-phase cDNA library.

Dynabeads™ mRNA Purification Kit (for mRNA purification from total RNA preps) (Invitrogen™)

The Dynabeads® mRNA Purification Kit rapidly isolates the mRNA transcriptome in typically 15 minutes, delivering pure, intact mRNA. The kit is designed to specifically target, capture, and purify mRNA molecules from total RNA preparations. Advantages of using the Dynabeads® mRNA Purification Kit:

• Obtain pure mRNA in typically 15 minutes
• Recover and enrich the transcriptome efficiently
• Prepare mRNA that is suitable for nearly every downstream application

Pure, rapid mRNA isolation
The Dynabeads® mRNA Purification Kit contains magnetic beads for the isolation of the mRNA transcriptome from any total eukaryotic RNA preparation (see figure). Dynabeads® magnetic beads are uniform in size and highly mobile in solution (see figure). This enables the bead capture surface to quickly and continuously interact with the entire total RNA sample during the mRNA capture phase. The ribosomal RNA and small RNA molecules (transfer RNA, microRNA, small nucleolar RNA, and small cytoplasmic RNA) do not bind to the beads and are discarded. Only polyadenylated RNA species (mRNA) are captured resulting in cleaner, more sensitive results (see figure). Within about 15 minutes, pure mRNA is isolated and ready for use in downstream applications.

A straightforward enrichment procedure
The Dynabeads® mRNA Purification Kit uses a robust affinity purification principle for the enrichment of polyadenylated mRNA. Superparamagnetic Dynabeads®, coupled to oligo-(dT)25, are first equilibrated with Binding Buffer, and then mixed with purified total RNA. The beads are then washed to remove contaminating RNA species, and then mRNA is eluted in as little as 5 µL of 10 mM Tris-HCl. The entire process is facilitated by the use of a neodymium magnet (purchased separately), which allows for the quick and efficient immobilization of the magnetic beads during buffer changes. RNA is suitable for all downstream molecular applications, including:

• Gene cloning
• cDNA synthesis, cDNA library construction
• RT-PCR, quantitative RT-PCR
• RPA (Ribonuclease Protection Assay)
• Subtractive hybridization
• Dot/slot hybridization
• Primer extension

Dynabeads™ mRNA DIRECT™ Purification Kit (Invitrogen™)

The Dynabeads® mRNA DIRECT™ Kit is designed for simple and rapid isolation of pure, intact polyadenylated (polyA) mRNA directly from the crude lysate of animal and plant cells and tissues. The isolated mRNA is suitable for use in all downstream applications. Advantages of the Dynabeads® mRNA DIRECT™ Kit:

Fast—15 minute procedure yields pure, intact mRNA
Highly pure mRNA isolation—best choice upstream of cDNA synthesis
Sensitive mRNA isolation—enables cDNA synthesis and cDNA library construction from ultra-small starting samples

System overview
The isolation protocol relies on base pairing between the polyA residues at the 3' end of most mRNA, and the oligo (dT)25 residues covalently coupled to the surface of the Dynabeads® . Other RNA species lacking a polyA tail will not hybridize to the beads and are readily washed away. Ribosomal RNA, DNA, proteins, and small RNA molecules (such as transfer RNA, micro RNA, and small nucleolar RNA) do not bind to the beads and are discarded. RNase inhibiting agents in the Lysis/Binding Buffer together with stringent hybridization and washing conditions ensure the isolation of pure, intact mRNA from crude samples rich in RNase, without the use of strong chaotropic agents. The mRNA purification beads specifically target and capture the mRNA transcriptome from an extremely wide variety of crude starting samples (see protocol). 1 mg of Dynabeads® Oligo (dT)25 beads (200 µL) binds up to 2 µg of mRNA. A typical mammalian cell contains about 10–30 pg of total RNA, from which 1–5% is mRNA.

Dynabeads™ Untouched™ Human B Cells Kit (Invitrogen™)

Use this kit to isolate pure and viable untouched B cells from PBMC by negative isolation. The kit depletes T cells, NK cells, monocytes, platelets, dendritic cells, granulocytes and erythrocytes. The negatively isolated human B cells are left in the sample and have not been in contact with the Dynabeads. The bead and antibody-free B cells are truly untouched, and in perfect shape for any functional assay and application.

• High purity, recovery and viability of human untouched B cells
• Easy to use and to scale up
• No columns required

An antibody mix towards the non-B cells is added to the sample and allowed to bind to the cells. Dynabeads® are added and will bind to the antibody-labeled cells during a short incubation. The bead-bound cells are quickly separated on a magnet and discarded. The remaining negatively isolated and untouched human B cells can be directly analyzed in a flow cytometer and used in any application.

Starting Sample:
PBMC

Dynabeads™ His-Tag Isolation and Pulldown (Invitrogen™)

This product is designed for his-tagged protein isolation. High selectivity for poly-histidine tags results in extremely low background levels. The new bead surface chemistry is Cobalt-based and offers exceptionally fast binding kinetics and a high yield per microliter of beads. The entire protocol (from cleared lysate to purified protein) takes only 20 minutes! The protocol includes instructions for making the required buffers as well as methods for optimizing buffer composition. Dynabeads® His-Tag Isolation & Pulldown replaces Dynabeads® TALON™. Dynabeads® His-Tag Isolation & Pulldown has a new optimized binding chemistry that offers significant advantages over Dynabeads® TALON™ which were coated in the TALON™ chemistry. This optimization results in a cleaner prep with higher yield of his-tag protein and extremely low background levels. Dynabeads His-Tag Isolation & Pulldown is designed for isolating His-tagged proteins from cleared lysates. Isolated his-tagged proteins can either be eluted from the beads or kept on the beads for use in bead-based downstream assays. Bead bound his-tagged proteins can be used in various downstream assays such as bait-prey pulldowns, etc. Note: This product contains Dynabeads and a protocol only, and does not include buffers. The technology used for the discontinued Dynabeads® TALON™ was licensed from Clontech. TALON™ is a registered trademark of Clontech Laboratories Inc., USA.

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.

Dynabeads™ Human T-Expander CD3/CD28 (Gibco™)

Dynabeads® Human T-Expander CD3/CD28 is intended for separation, activation and expansion of human T cells. Dynabeads® Human T-Expander CD3/CD28 is the research grade version of Dynabeads® CD3/CD28 CTS™. Containing the same amount and ratio of antibodies from the same clones as Dynabeads® CD3/CD28 CTS™, Dynabeads® Human T-Expander CD3/CD28 are intended for small scale pre-clinical research. Dynabeads® Human T-Expander CD3/CD28 offers a simple method for activation and expansion of T-cells that does not require antigen presenting cells or antigen. By combining anti-CD3 and anti-CD28 antibodies on Dynabeads®, the beads provide both the primary and co-stimulatory signals that are required for activation and expansion of T cells.

Applications:
Dynabeads® Human T-Expander CD3/CD28 is the research grade version of Dynabeads® CD3/CD28 CTS™. The Dynabeads® CD3/CD28 CTS™ technology has been successfully used in ex vivo separation, activation and expansion of T-cells in a number of studies, including studies on HIV infection and cancer. Other potential applications for this novel technology are treatment of autoimmune and infectious diseases and prevention of complications associated with chemotherapy and allogeneic transplantation.