Invitrogen™ DynaGreen™ Protein A Magnetic Beads

Catalog No.	Quantity
80102G	3 mL, 120 Reactions
80101G	0.5 mL, 20 Reactions

Catalog No. 80102G Supplier Invitrogen™ Supplier No. 80102G

Description

DynaGreen Protein A Magnetic Beads are intended for use in immunoprecipitation (IP) applications while also being an environmentally sustainable choice.

DynaGreen Protein A Magnetic Beads are intended for use in immunoprecipitation (IP) applications while also being an environmentally sustainable choice. They are microplastic-free submicron magnetic beads that provide reproducible isolation of target proteins in a simple and efficient workflow, which can be performed manually or automated on KingFisher™ instruments.

Benefits of DynaGreen Protein A Magnetic Beads include:

• High performance—reproducible IP with high-yield and low non-specific binding
• Reduced environmental impact—non-microplastic bead core and holistic sustainable product design
• Flexible—direct and indirect immunoprecipitation helps ensure isolation of non-abundant proteins
• Automation ready—easy automation for high-throughput needs on the Kingfisher Sample Purification Systems

Simple, gentle, and efficient manual separation

DynaGreen Protein A Magnetic Beads are microplastic-free superparamagnetic beads with recombinant protein A (∼45 kDa) covalently coupled to the surface. They were developed for convenient IP of target proteins when the IP antibody interacts strongly with Protein A (see subclass specificity in the manual). The submicron bead size (∼250 nm) provides low sedimentation and a large target capture surface area. The method is a simple bind-incubate-elute protocol with no pre-clearing required. This results in an efficient high-yield isolation of target protein by direct or indirect immunoprecipitation in less than 80 minutes.

The magnetic separation technology used by DynaGreen Protein A Magnetic Beads is rapid and gentle, thereby reducing physical stress to the target proteins. Furthermore, the non-porous nature of the beads lowers the risk of antibodies being trapped and becoming inaccessible. This helps to minimize the amount of antibody required to produce accurate, and reproducible results. The yield and purity of the isolated target protein are highly compatible for use in western blotting and mass spectrometry. The product manual below includes a list of required reagents, materials, and details of compatible buffer recipes.

Rapid and reproducible automated workflow

The DynaGreen magnetic beads are ideal for high-throughput enrichment. This process can be automated using any of our KingFisher sample purification systems or liquid handlers.

The automated protocol can be performed in just 40 minutes with up to 96 samples per run, which helps reduce hands-on time while maintaining the same reproducible high target yield and low non-specific binding compared to the manual protocol.

A certified choice for environmentally sustainable research

Microplastic-free DynaGreen Protein A Magnetic Beads are designed for both high performance in immunoprecipitation workflows and lower environmental impact. They provide exceptional purity and high yield of target proteins compared to other magnetic beads in both direct and indirect immunoprecipitation workflows. At the same time, the product boasts an extensive list of third-party ACT-label-certified sustainable product design elements by incorporating principles of green chemistry and green engineering.

Choosing DynaGreen Protein A Magnetic Beads can help reduce the environmental impact of research without compromising on results.

Learn more about our DynaGreen magnetic beads and KingFisher Sample Purification Systems:

• Alternative sustainable products: DynaGreen Protein A/G (Cat. No. 80106G) and DynaGreen Capture-Select anti-FC (multispecies) (Cat. No. 80107G)
• DynaGreen magnetic beads sustainability
• DynaMag magnets for magnetic bead separations
• KingFisher automation protocols
• KingFisher Flex instrument video

Specifications

• Concentration: 20 mg/mL
• Description: Recombinant Protein A covalently bound to DynaGreen magnetic beads
• Material: Iron Oxide
• Content And Storage: DynaGreen Protein A Magnetic Beads are supplied in phosphate buffer, pH 7.4, with a biodegradable surfactant and a preservative. Store at 2–8°C.
• Diameter (Metric): 250 nm
• For Use With (Equipment): KingFisher™ Sample Purification Systems, DynaMag™ Magnets
• High-throughput Compatibility: High-throughput Compatible
• Shipping Condition: Ambient temperature
• Product Line: DynaGreen
• Quantity: 3 mL, 120 Reactions
• For Use With (Application): Immunoprecipitation
• Target: Proteins
• Type: Protein A Magnetic Bead
• Ligand Type: Recombinant Protein A
• Sufficient For: 120 Tests
• Color: Brown
• Purity or Quality Grade: For Research Use Only
• Certifications/Compliance: ISO9001 and ISO13485

Frequently Asked Questions (FAQs)

What can I do if the antibody from the DynaGreen magnetic beads is coming off the beads during the elution step?

The antibody will come off the beads during elution if the antibody is not cross-linked to the beads. The antibody is attached to the Protein A/G through affinity binding, thus when you break one of the affinity bindings, the other affinity bindings will break as well.

Do you have any tips to offer if we are experiencing non-specific protein binding to DynaGreen magnetic beads or proteins trapped in the protein-antibody-antigen immunocomplex?

Here are some tips:
- Perform a pre-clearing procedure before doing immunoprecipitation (IP) by adding the beads to the sample without the antibody bound. After this pre-clearing step, continue with the IP protocol using the antibody-bound beads.
- Increase the washing time and increase the number of washing steps after IP.
- Increase the stringency of the washes, e.g., wash with a buffer containing a higher concentration of detergent or salt.
- Try the indirect IP technique (add antibody to the sample first, and then add the beads), change to another DynaGreen product (e.g., switch from DynaGreen Protein A/G Magnetic Beads to DynaGreen CaptureSelect Anti IgG-Fc Magnetic Beads), or try Dynabeads Protein A or Dynabeads Protein G for Immunoprecipitation.

When using DynaGreen magnetic beads, how can we avoid sample degradation by proteases?

We recommend including additional protease inhibitors in the lysis and wash buffers and keeping the sample cold at all times.

How can I improve the elution of the target from DynaGreen magnetic beads using a low pH buffer?

The addition of a small amount of detergent (eg., 0.05% of Tween 20) in the wash buffer, before elution, can improve the elution of the target. Also, it is important to fully remove the wash buffer before adding the elution buffer, in order to maintain the low pH conditions required for elution.

Are DynaGreen magnetic beads sterile?

Per definition, DynaGreen magnetic beads are not sterile, but all the buffers used in the manufacturing process are autoclaved and a biocide called Acticide is added to the product to help keep it free of any type of growth.

Can I freeze DynaGreen magnetic beads?

In general, we do not recommend freezing DynaGreen magnetic beads as freezing can affect the functionality of the beads.

Can I sonicate DynaGreen magnetic beads?

In general, short sonication is a good way to reduce aggregation of DynaGreen magnetic beads and ensure optimal homogenous conditions at the time of ligand addition when coating the beads. When the target is bound to the beads, one needs to be more careful with sonication as the binding might break.

What are the main differences between Dynabeads magnetic beads and DynaGreen magnetic beads?

DynaGreen magnetic beads differ from Dynabeads magnetic beads in bead size, chemistry, production time, and environmental sustainability profile; DynaGreen magnetic beads are submicron particles with an iron oxide core. They are produced in a few weeks with a more sustainable manufacturing process while Dynabeads magnetic beads are micron-sized particles, produced in 12-16 weeks with a magnetized polymer core.

Are DynaGreen magnetic beads functional if they have aggregated?

Aggregation and change of color is expected and does not affect the functionality of DynaGreen magnetic beads. If aggregation is observed upon storage of the beads, a few minutes of sonication in a water bath will restore the homogeneity of the bead suspension.

What is the shelf life of DynaGreen magnetic beads?

DynaGreen magnetic beads are supplied with a 12 month shelf life.

Is blocking necessary when using DynaGreen magnetic beads?

When using the buffers described in the manual, there is no need for further blocking. However, this might be different in other workflows and with other sample types. Also, if there is any non-specific binding, different blockers should be explored.

Can I reuse DynaGreen magnetic beads and do you have a protocol available for reuse?

We have not tested reuse of DynaGreen magnetic beads, but if the elution is gentle, in theory, it should be possible. Normally we do not recommend reusing products, but upon dissociation of antibody from the protein using various protocols (low pH, chaotropes), the beads should be reusable.

Do you have an antibody crosslinking protocol for DynaGreen magnetic beads?

For DynaGreen magnetic beads, we recommend using a lower concentration of BS3 in the conjugation buffer than what is recommended for Dynabeads magnetic beads (from 1.7 mM to 2.5 mM instead of 5 mM). Also, we recommend adding Tween 20 to the conjugation buffer to a final concentration of 0.05% in order to improve the performance of the crosslinker and of the beads. We recommend using DynaGreen Protein A or DynaGreen Protein A/G magnetic beads for this workflow.

What is the stability of DynaGreen magnetic beads at elevated temperatures?

Our stability studies have shown that prolonged exposure of DynaGreen magnetic beads to elevated temperatures reduces the functionality of the beads. However, beads still retained functionality after three weeks at +50 degrees C.

What is the concentration of DynaGreen magnetic beads in number of beads per mL?

DynaGreen magnetic beads are submicron particles. Since the beads are so small, there isn't a good way to count them, and the bead concentration is expressed as mg/mL.

Why does the immunoprecipitation (IP) protocol for DynaGreen magnetic beads have a longer incubation time (80 min) than the one for Dynabeads magnetic beads (40 min incubation time)?

DynaGreen magnetic beads have excellent magnetic properties. The reason for the slightly longer manual immunoprecipitation (IP) time is purely based on selection of an incubation time that gave the best target recovery in our workflow. We recommend optimizing your workflow with regards to both incubation time and amounts of reagents added. Shorter incubation time (40 min protocol) has been tested successfully in our workflow and in fact, the KingFisher IP protocol for DynaGreen magnetic beads is 40 min (same as that for Dynabeads magnetic beads).

What is the compatibility of DynaGreen magnetic beads with chelators, reducing agents, and organic solvents?

Chelators and stronger reducing agents like thiols will tend to remove the iron from the bead core. Short exposure to up to 0.1 M reducing agents and chelators should, in theory, be tolerated.

Are there any pH limits with DynaGreen magnetic beads for washing or elution steps?

High pH (>9) might have a negative effect on the bead structure, however, short incubation times at high pH may be acceptable. Low pH (2.8) has successfully been used for gentle elution of antibodies and target from DynaGreen magnetic beads, in our workflows.

Do you offer a DynaGreen-based product that is equivalent to the Dynabeads Antibody Coupling Kit?

Currently, we do not offer such a product, but it is being considered for our future offering on this platform. We have successfully coupled antibodies to these beads as well as other ligands.

How do you score sustainability for DynaGreen magnetic beads?

My Green Lab reviews products from manufacturing, user impact, and end of product life to qualify them for the ACT (Accountability, Consistency, and Transparency) Environmental Factor Label. Currently, our DynaGreen magnetic beads are the only beads with such a label, so a direct comparison with competitors is not possible, but we believe that this transparency will provide confidence to the customer (https://act.mygreenlab.org/).

Which magnets can I use with DynaGreen magnetic beads in 96 well plates?

The DynaGreen magnetic beads have excellent magnetic properties, so they should work well with any magnet. DynaMag magnets of all sizes have been used in-house; we routinely use DynaMag-96 Side-Skirted Magnet, as well as DynaMag-2 and DynaMag-50 tube magnets.

Can you comment on the non-specific binding when using DynaGreen magnetic beads to isolate exosomes?

When DynaGreen magnetic beads are used for exosome isolation, the non-specific binding is low, especially when DynaGreen CaptureSelect anti IgG-Fc beads are used. These beads have been used successfully for isolation of exosomes from buffer, plasma, and urine.

Can you provide specific information about the protein ligands used in the DynaGreen magnetic beads?

The proteins in the DynaGreen magnetic beads are recombinant and animal origin-free. Protein A and Protein A/G are produced in E. coli. CaptureSelect anti-IgG Fc is produced in yeast (S. cerevisiae) and is a camelid-derived single domain (VHH), 14 kDa antibody fragment.

What is the binding capacity of DynaGreen magnetic beads?

The binding capacity of DynaGreen magnetic beads is typically between 12 and 20 µg human gammagobulin per mg beads. Note that the beads have been developed with immunoprecipitation (IP) performance and cost in mind, and not to maximize the binding capacity.

When using DynaGreen magnetic beads, how much should I dilute the lysate with RIPA buffer?

We routinely use RIPA Lysis and Extraction Buffer for lysing cells (Cat. No. 89900, 89901). No other dilutions of RIPA buffer have been tested in-house. Other common lysis buffers have been tested successfully by customers.

When using detergent with DynaGreen magnetic beads, do you have a specific workflow for mass spectrometry (MS) application to avoid carryover of detergent?

For mass spectrometry (MS) application, we recommend the following:
- Increasing the amount of starting material, DynaGreen magnetic beads, and antibody.
- Increasing the binding time of the antibody to the beads to 1 hour. Please note that optimization may be required for each antibody and target antigen.
- Increasing the number of washes at the end of the procedure and washing with more stringent reagents to decrease non-specific background. We recommend using PBS without Tween 20 in the last 2 washes to remove detergent.
- Beads and protocol-recommended buffers are compatible with on-bead peptide digestion.
Note: Due to their small size, the DynaGreen magnetic beads are prone to packing together once the detergent is removed, and less easy to disperse, so be mindful to keep the beads collected at the bottom of the tube.

For Research Use Only. Not for use in diagnostic procedures.