DynaGreen™ CaptureSelect™ Anti-IgG-Fc (Multi-Species) Magnetic Beads, 0.5 mL, 20 rxns - FAQs

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.

Find additional tips, troubleshooting help, and resources within our Protein Immunoprecipitation (IP), Co-Immunoprecipitation (Co-IP), and Pulldown Support Center.

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.

Find additional tips, troubleshooting help, and resources within our Protein Immunoprecipitation (IP), Co-Immunoprecipitation (Co-IP), and Pulldown Support Center.

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

Find additional tips, troubleshooting help, and resources within our Protein Immunoprecipitation (IP), Co-Immunoprecipitation (Co-IP), and Pulldown Support Center.

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.

Find additional tips, troubleshooting help, and resources within our Protein Immunoprecipitation (IP), Co-Immunoprecipitation (Co-IP), and Pulldown Support Center.

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.

Find additional tips, troubleshooting help, and resources within our Protein Immunoprecipitation (IP), Co-Immunoprecipitation (Co-IP), and Pulldown Support Center.

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

Find additional tips, troubleshooting help, and resources within our Protein Immunoprecipitation (IP), Co-Immunoprecipitation (Co-IP), and Pulldown Support Center.

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.

Find additional tips, troubleshooting help, and resources within our Protein Immunoprecipitation (IP), Co-Immunoprecipitation (Co-IP), and Pulldown Support Center.

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.

Find additional tips, troubleshooting help, and resources within our Protein Immunoprecipitation (IP), Co-Immunoprecipitation (Co-IP), and Pulldown Support Center.

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.

Find additional tips, troubleshooting help, and resources within our Protein Immunoprecipitation (IP), Co-Immunoprecipitation (Co-IP), and Pulldown Support Center.

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

Find additional tips, troubleshooting help, and resources within our Protein Immunoprecipitation (IP), Co-Immunoprecipitation (Co-IP), and Pulldown Support Center.

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.

Find additional tips, troubleshooting help, and resources within our Protein Immunoprecipitation (IP), Co-Immunoprecipitation (Co-IP), and Pulldown Support Center.

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.

Find additional tips, troubleshooting help, and resources within our Protein Immunoprecipitation (IP), Co-Immunoprecipitation (Co-IP), and Pulldown Support Center.

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.

Find additional tips, troubleshooting help, and resources within our Protein Immunoprecipitation (IP), Co-Immunoprecipitation (Co-IP), and Pulldown Support Center.

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.

Find additional tips, troubleshooting help, and resources within our Protein Immunoprecipitation (IP), Co-Immunoprecipitation (Co-IP), and Pulldown Support Center.

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.

Find additional tips, troubleshooting help, and resources within our Protein Immunoprecipitation (IP), Co-Immunoprecipitation (Co-IP), and Pulldown Support Center.

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).

Find additional tips, troubleshooting help, and resources within our Protein Immunoprecipitation (IP), Co-Immunoprecipitation (Co-IP), and Pulldown Support Center.

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.

Find additional tips, troubleshooting help, and resources within our Protein Immunoprecipitation (IP), Co-Immunoprecipitation (Co-IP), and Pulldown Support Center.

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.

Find additional tips, troubleshooting help, and resources within our Protein Immunoprecipitation (IP), Co-Immunoprecipitation (Co-IP), and Pulldown Support Center.

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.

Find additional tips, troubleshooting help, and resources within our Protein Immunoprecipitation (IP), Co-Immunoprecipitation (Co-IP), and Pulldown Support Center.

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/).

Find additional tips, troubleshooting help, and resources within our Protein Immunoprecipitation (IP), Co-Immunoprecipitation (Co-IP), and Pulldown Support Center.

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.

Find additional tips, troubleshooting help, and resources within our Protein Immunoprecipitation (IP), Co-Immunoprecipitation (Co-IP), and Pulldown Support Center.

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.

Find additional tips, troubleshooting help, and resources within our Protein Immunoprecipitation (IP), Co-Immunoprecipitation (Co-IP), and Pulldown Support Center.

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.

Find additional tips, troubleshooting help, and resources within our Protein Immunoprecipitation (IP), Co-Immunoprecipitation (Co-IP), and Pulldown Support Center.

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.

Find additional tips, troubleshooting help, and resources within our Protein Immunoprecipitation (IP), Co-Immunoprecipitation (Co-IP), and Pulldown Support Center.

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.

Find additional tips, troubleshooting help, and resources within our Protein Immunoprecipitation (IP), Co-Immunoprecipitation (Co-IP), and Pulldown Support Center.

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.

Find additional tips, troubleshooting help, and resources within our Protein Immunoprecipitation (IP), Co-Immunoprecipitation (Co-IP), and Pulldown Support Center.