PureCube Amine Activated MagBeads available as off-the-shelf items. Our careful in-house production provides low lot-to-lot variation, making activated MagBeads also suitable for commercial production. PureCube Amine Activated MagBeads are provided as a 25% suspension. PureCube Amine Activated MagBeads carry an amine functionality. Proteins can bind covalently via free carboxy groups after activation with EDC/NHS.
PureCube Amine Activated MagBeads provide:
||Coupling of biomolecules via EDC for affinity purification via free Carboxy groups
||Covalent coating of biomolecules via EDC to free carboxy groups
||Delivered as a 25 % suspension
|amino Group density
|| higher than 15 µmol/ml
||Activated Amine groups
- Coupling buffer I (PBS): 150 mM Na phosphate, 100 mM NaCl, pH 7.2
- Coupling buffer II: 0.1 M MES, 150 mM NaCl, pH 4.7
- Wash buffer I (PBS): 150 mM Na phosphate, 100 mM NaCl, pH 7.2
- Wash buffer II: 250 mM NaCl
- Storage buffer I: 20 mM sodium acetate pH 6,5, 20% Ethanol
- Storage buffer II: 100 mM sodium hydrogen carbonate, 0.02% sodium azide, pH 7.5
- EDC ((1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide). It is highly recommended to open EDC bottles under protective gas and to let the EDC equilibrate to room temperature before opening. Use EDC directly after withdrawal from storage vessel! Important: If you are not sure if the EDC has been in contact with humidity, discard the material and use fresh EDC.
- Magnetic holder for microcentrifuge tubes (for separation of magnetic beads)
- Microcentrifuge tubes (2 mL)
- Centrifuge for 15 mL tubes
- Centrifuge tubes (15 mL)
- End-over-end mixer or thermomixer
|A.||Amine coupling protocol:|
- Dispense 800 µl of the 25% magbead suspension into a 2 ml microtube and place the tube in a MagBead seperator
- Remove the supernatant and wash the magbeads three times with 1 ml PBS each.
- Resuspend 400 µg to 2 mg protein in 200 µl PBS, add them to the suspension and mix by vortexing.
- Incubate the reaction mixture for 5-10 minutes at 4°C on an end-over-end shaker.
- Depending on the stability of your protein, incubate for 2h at 4°C or 1h at room temperature on an end-over-end shaker.
- In some cases, the binding capacity can be raised by a second addition of 4 mg EDC and additional 1-2h incubation.
- Wash the mag beads five times with 1 ml PBS and once with 1 ml double distilled water. Resuspend the magbeads in 800 µl storage buffer. Keep at 4°C until further use.