Can I store the antibody/ligand-coated Dynabeads magnetic beads?
Dynabeads magnetic beads coated with antibody/ligand may be stored at 2-8 degrees C without loss of antigen binding capacity. For long-term storage, a final concentration of 0.02% NaN3 may be added to the antibody-coupled beads in a physiological buffer. Please note that not all coupled antibodies retain their function in long term storage. Verify your coupled antibody stability by testing in small scale. After storage, coated Dynabeads magnetic beads should be washed once in PBS/BSA for 5 min before use.
Find additional tips, troubleshooting help, and resources within ourProtein Immunoprecipitation (IP), Co-Immunoprecipitation (Co-IP), and Pulldown Support Center.
How can I isolate cells using secondary-coated Dynabeads magnetic beads?
The secondary-coated Dynabeads magnetic beads can be coupled to a primary antibody by a direct or an indirect approach.
Direct approach: The Dynabeads magnetic beads are first coupled with your primary antibody and then used for isolating your target cell type.
Indirect approach: The cells are first incubated with your primary antibody(ies). The Dynabeads magnetic beads are then added to the antibody-coated target cells.
Secondary-coated Dynabeads magnetic beads can be used in several cell isolation approaches:
Cell depletion--using an antibody to target the unwanted cell type and a secondary-coated Dynabeads magnetic beads product.
Negative cell isolation--using a cocktail of antibodies to target all unwanted cell types and a secondary-coated Dynabeads magnetic beads product (using the indirect approach).
Positive cell isolation without detachment--using an antibody to target the wanted cell type and a secondary-coated Dynabeads magnetic beads product.
Positive cell isolation with detachment--using an antibody to target the wanted cell type and a CELLection Dynabeads magnetic beads product.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
If Thermo Fisher Scientific does not have a primary-coated Dynabeads magnetic beads product for isolating my target cell type, which alternative Dynabead magnetic beads product can I use instead?
You may use one of our secondary-coated, surface-activated, or streptavidin-coated Dynabeads magnetic beads, and coat it with a primary antibody to target your cell type.
The Dynabeads magnetic beads product you choose will depend on the primary antibody available for cell targeting and the downstream application for the isolated cells:
-For primary antibodies made in mouse, use the CELLection magnetic beads Pan Mouse IgG Kit, Dynabeads magnetic beads Goat Anti-Mouse IgG, Dynabeads magnetic beads Pan Mouse IgG, Dynabeads magnetic beads Rat Anti-Mouse IgM, Dynabeads magnetic beads Rat Anti-Mouse IgM, or Dynabeads magnetic beads Sheep-Anti Mouse IgG
-For primary antibodies made in rat, use the Dynabeads magnetic beads Sheep Anti-Rat IgG
-For primary antibodies made in rabbit, use the Dynabeads magnetic beads M-280 Sheep Anti-Rabbit IgG
-For primary antibodies made in any species, use the CELLection magnetic beads Biotin Binder Kit, Dynabeads magnetic beads Biotin Binder, Dynabeads magnetic beads FlowComp Flexi, Dynabeads magnetic beads M-450 Epoxy, or Dynabeads magnetic beads M-450 Tosylactivated
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
Can a cocktail of primary antibodies be added to a cell suspension in order to pull out several target cell populations simultaneously, using one secondary-coated Dynabeads magnetic beads product?
Yes, a cocktail of primary antibodies can be added to a cell suspension in order to pull out several target cell populations with one secondary-coated Dynabeads magnetic beads product.
The Dynabeads magnetic beads Pan Mouse IgG (110.41; 110.42) works very well with a cocktail of mouse IgGs for the simultaneous capture of multiple cell types. it is recommended that you use an indirect technique with antibody cocktails (add all Ab to cells, wash off excess Ab, then add beads to capture Ab-coated cells).
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
When do I use the direct or indirect isolation techniques?
The indirect technique is chosen when the antigen targeted by the primary antibody is expressed in low density on the target cell surface. This is due to the fact that free antibodies will find their target antigen more easily than antibodies linked to the Dynabeads magnetic beads. Also when using the indirect technique, an excess of free antibody can be added to the system, allowing ample opportunity for monoclonals to find the target antigen. Finally, an indirect technique can be useful when a cocktail of monoclonal antibodies is used to deplete unwanted cells during negative isolation of a cell type. This is because antibodies against all unwanted cell types can be added at once to the starting cell population, provided the antibodies are from one species. The antibody-coated cells can then be targeted with secondary-coated Dynabeads magnetic beads.
The direct technique is chosen when a limiting amount of monoclonal antibody is needed for targeting the cells of interest during positive isolation or depletion (e.g., when the target antigen is present at high density). It can also help when the possibility of interaction from the secondary antibody needs to be avoided, or if a stock preparation of primary coated Dynabeads magnetic beads is desired. Additionally the direct technique can be used when you do not want to cover all antigen sites with antibody (e.g., when you want to analyze the isolated cells by flow cytometry).
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.