Cat# 21001
Product Description
MagVigen™ – NH2 surface nanoparticles provide you with the flexibility of coupling to various molecules through simple bioconjugation reactions. The resulting MagVigen™ bioconjugates could be exploited to achieve highly specific binding for cell isolation, protein, DNA/RNA purification or immunoprecipitation assays. Examples of biomolecules that could be covalently bound to MagVigen™ surfaces include primary antibody, protein/peptide, DNA/RNA or other ligands.
Advantages of MagVigen™ magnetic nanoparticles
- Magnetically responsive to a magnet, easy for bio-conjugation and purification
- Smaller nanoparticle size, higher binding capacity, longer settling time, compatible to automation and high throughput workflow
- Optimal surface chemistry, low non-specific binding
- Consistent, high quality results
Product and Related Product Contents
- MagVigen™ – NH2 surface nanoparticles (Cat# 21001) are provided in phosphate buffered saline (PBS), pH 7.4. Each vial contains 1 ml of solution with a particle concentration of 4 mg/ml.
- Washing Buffer (10X), Cat# A20001.
- Magnet, Cat# A20003.
All materials except the magnet should be stored at 4°C for up to 1 year.
Protocol
I.This protocol provides a general guidance for conjugation of biomolecules to MagVigen™-NH2 surface through EDC/NHS crosslinking of carboxylates with primary amines on nanoparticle surfaces. Please adjust the amount of reagents for specific application.
- Determine needed surface coverage of biomolecules per nanoparticle.
- Dissolve the protein in a buffer of 0.1M MES (2- [morpholino]ethanesulfonic acid) and 0.5M NaCl at pH 6.0, for example, prepare 0.2 ml of protein at 10 mg/ml.
- Add 0.08 mg of EDC to 0.2 ml of the above protein, based on a 50kDa protein, 10 fold excess of EDC (1-ethyl-3-[3- dimethylaminopropyl] carbodiimide) to protein quantity is applied.
- Add 0.12mg of NHS (N-hydroxysuccinimide) or 0.22mg of Sulfo-NHS (N-hydroxysulfosuccinimide) to the reaction (final concentration 5 mM).
- Mix the solution well and react for 15 min at room temperature.
- Gently vortex MagVigen™-NH2, then separate the nanoparticles from the solution by placing the magnet on the side of the tube for 1-2 min and remove the supernatant carefully (with magnet still on the side).
- Remove the magnet and disperse the pellet in 1ml of 1x Washing Buffer. Repeat step 6, wash once. Then re- disperse MagVigen™-NH2 in 1 ml of 1x Washing Buffer.
- Mix the EDC/NHS activated protein solutions with MagVigen™-NH2 solution; react for 2 hours at room temperature.
- Separate the nanoparticles from the solution by placing the magnet on the side of the tube for 1-2 min and remove the supernatant carefully (with magnet still on the side). Wash 2 times with PBS or other buffer solution. Remove non- magnetically captured solution.
- Resuspend washed MagVigen™-protein conjugates into preferred buffer, ready to use.
Note: The general range is about 0.1 -1 mg proteins (50kDa) per mg of MagVigen™.
Note: A clear precipitate containing dark brown colored nanoparticles should become visible on the side of the micro-centrifuge tube.
II. This protocol provides a general guidance for conjugation of biomolecules to MagVigen™-NH2 surface through sulfo-SMCC (Succinimidyl trans- 4(maleimidylmethyl) cyclohexane-1-carboxylate) based crosslinker utilizing -SH group of the biomolecule. Please adjust the amount of reagents for specific application.
- Determine needed surface coverage of biomolecules per nanoparticle.
- Gently vortex MagVigen™-NH2, then separate the™nanoparticles from the solution by placing the magnet on the side of the tube for 1-2 min and remove the supernatant carefully (with magnet still on the side).
- Remove the magnet and disperse the pellet in 1ml of 1x Washing Buffer. Repeat step 2, wash once. Then re- disperse MagVigen™-NH2 in 1 ml of 1x Washing Buffer.
- Add Sulfo-SMCC into 200 μl PBS buffer, then mix with above MagVigen™-NH2 solution, Incubate for 40-60 min.™The ratio of Sulfo-SMCC to MagVigen™ is at 0.05 mg of Sulfo-SMCC per 1 mg of MagVigen™.
- Separate the Sulfo-SMCC activated nanoparticles from the solution by placing the magnet on the side of the tube for 1-2 min, and then remove the supernatant carefully (with magnet still on the side). Then wash twice using 1x washing buffer.
- Dissolve desired amount of proteins into 1x PBS solution.
- Mix the protein solution with the Sulfo-SMCC activated nanoparticle solution, incubate for 2 hours. For maximal binding, overnight incubation could be applied.
- Separate the MagVigen™-protein conjugates from the solution by placing the magnet on the side of the tube for 1-2 min and remove the supernatant carefully (with magnet still on the side). Wash twice using 1x washing buffer or other preferred buffer.
- Resuspend washed MagVigen™-protein conjugates into preferred buffer, ready to use.
Note: The general range is about 0.1 -1 mg proteins (50kDa) per mg of MagVigen™.