Ideal for Immunoprecipitation of protein complexes:
Low background and covalent linking of antibodies to the bead surface make Dynabeads Tosylactivated an excellent choice for Immunoprecipitation of proteins and protein complexes (Co-Immunoprecipitation, Co-IP). Gentle and rapid magnetic concentration of the beads and short incubation times (possible due to fast surface-based binding kinetics) make Dynabeads Tosylactivated an excellent choice for the immunoprecipitation of highly-labile or and/or transient (short-lived) protein complexes.
Bead Surface Characteristics for Dynabeads Tosylactivated:
- p-toluene-sulfonyl (Tosyl) groups
- Hydrophobic, pH Neutral
- Covalent binding by primary amine (NH2) or sulphydryl (SH) groups
- 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
- 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 Tosylactivated. Ligands commonly coupled to Dynabeads Tosylactivated include peptides and proteins (e.g. antibodies for Immunoprecipitation or Co-Immunoprecipitation). Coupling occurs at neutral to high pH and at 37°C. We recommend coupling at pH 8.5-9.5, but for pH labile ligands, coupling can be performed in an alternative buffer at pH 7.4.
Upon completion of the ligand coupling step, the actual Dynabeads Tosylactivated surface coating will be rendered inert, resulting in low non-specific binding.
Binding capacity per milligram beads:
Varies depending on ligand, (e.g. 14-19 µg IgG)
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.