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