CELLection™ Biotin Binder Kit

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.

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.

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.

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.

When a secondary-coated Dynabeads magnetic beads product is used for negative isolation or depletion of cells you can choose between the following techniques.

Direct technique
-Add primary IgG antibody to the secondary-coated Dynabeads magnetic beads with specificity to the species of your primary IgG antibody
-Add the resulting primary coated Dynabeads magnetic beads to cells for capture and separation

Benefits:
-You can make up a stock of beads
-Use less primary antibody
-Save time, fewer steps and is easier to optimize than the indirect procedure
-Lower background, additional steps increase the chance of nonspecific signal
-Lower cost, since direct procedures require much lower amounts of tag specific antibodies than indirect procedures

OR

Indirect technique
-Add primary IgG antibody to the cells
-Wash cells to remove excess, unbound antibody
-Add secondary-coated Dynabeads magnetic beads with specificity to the species of your primary IgG antibody to capture primary antibody coated cells

Benefits:
-You can use a cocktail of antibodies to coat different cell types i.e., a negative isolation approach
-Can be used when target antigen expression is low for more efficient binding of antibody to cells.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.