Ideal for Immunoprecipitation of protein complexes:
Ultra-low background and covalent linking of antibodies to the bead surface make Dynabeads Epoxy ideal for Immunoprecipitation of proteins and protein complexes (Co-Immunoprecipitation, Co-IP). Gentle yet rapid magnetic concentration of the beads and short incubation times (possible due to fast surface-based binding kinetics) make Dynabeads Epoxy the best choice for the immunoprecipitation or protein complexes, especially when those complexes are highly-labile or transient (short-lived).
Ultra rapid protocols:
- Identify members of protein complexes in minutes rather than hours
- Temporal resolution short enough to identify transient and labile complexes
- Identify binding partners that cannot be identified with longer protocols
- Rapid protocols further reduce the already ultra-low background binding
- Stronger signal to noise-ratios
Applications:
- IP of proteins and protein complexes
- Couple functional enzymes to bead surface for downstream assays
- Couple peptides to bead surface to identify binders
Purify peptides, proteins and enzymes that are:
- Transiently stable
- Structurally intact
- Temperature labile
- In their native conformation and functional
Coupling procedure outline:
Covalent coupling is performed overnight by incubating the desired ligand with the Dynabeads Epoxy. Ligands commonly coupled to Dynabeads Epoxy include peptides and proteins (e.g. antibodies for Immunoprecipitation or Co-Immunoprecipitation). Coupling occurs at neutral pH and high salt concentration. The coupling reaction will occur in a broad temperature range making it possible to choose the temperature that is best suited to the stability characteristics of your ligand.
Upon completion of the ligand coupling step, the active coupling chemistry on the Dynabeads Epoxy surface coating will be rendered inactive, such that no further proteins will be covalently coupled to the bead surface. The nature of the epoxy groups, once the coupling reaction is completed, is such that these beads will exhibit extremely low non-specific binding. The hydrophilic and pH neutral bead surface also enhances the extremely low non-specific binding characteristics of these beads.
About Dynabeads:
Dynabeads are non-porous monodisperse superparamagnetic beads. They are highly mobile in solution enabling ligands coupled to the beads to continuously interact with the entire sample volume. The superparamagnetic beads are pulled to the tube walls by transferring the tube to a rack containing a strong magnetic field. Strong magnetic fields quickly pull the beads to the tube wall, allowing for easy and complete removal of the supernatant by pipette. Washing steps are performed similarly.
Bead Surface Characteristics for Dynabeads Epoxy:
- Coated in glycidyl ether (Epoxy) groups
- Hydrophilic, pH Neutral
- Covalent binding by primary amine (NH2) or sulphydryl (SH) groups
Binding capacity per milligram beads:
Varies depending on ligand, (e.g. 5-10 µg IgG)
