Cat# 21005C / K21005C
Product Description
MagVigen™-Streptavidin magnetic nanoparticles can universally bind to any biotinylated biomolecules (e.g., antibody, protein, peptide, DNA) through high affinity interaction between streptavidin and biotin. The MagVigen™-Streptavidin-biotin-biomolecule complex can be easily separated from unbound biotin-biomolecule using a magnetic rack (Cat#A20006). This provides a quick and neat way to tag biomolecules with magnetic nanoparticles. The purified nanoparticle-biomolecule complex can be used in a variety of downstream bio-separation processes (e.g., protein purification, immunoprecipitation, cell isolation or depletion, and molecular detection.)
MagVigen™- Streptavidin are ideally used together with biotin- antibody for isolation of cells (e.g., CTCs, stem cells) from a mixture of cell population obtained from tissues or organs. The isolated cells are viable and can be further cultured or used for downstream molecular analysis such as mRNA isolation and RT-PCR. Cell separation with MagVigen™ nanoparticles eliminates the use of columns, so cells are not exposed to the mechanical stress from passing through the column matrix. Magnetically separated cells are highly purified and retain their viability, ideal for downstream applications.
Advantages of MagVigen™ – Streptavidin for Molecular and Cellular Enrichment
- Easy and quick to make nanoparticle-primary antibody onjugates
- Consistent, high-quality results
- High binding capacity, 25-50 µg biotin-antibody per mg of nanoparticles
- High biocompatibility
- Low non-specific binding
Product Contents
- Cat# 21005C: MagVigen™- Streptavidin are provided in phosphate buffered saline (PBS) containing 0.05% NaN3, 0.1% BSA. pH 7.4. Each vial contains 1 ml of solution with particle concentration of 2 mg/ml, which is enough for binding 50 µg of biotin-antibody.
Nanoparticle size: 200-500 nm measured using Dynamic Light Scattering.
Polydispersity index < 0.2.
Capacity: 50μg biotin-antibody/ml of nanoparticles
Cat# K21005C further includes:
- Wash Buffer
- Antibody Binding Buffer
All materials except the magnet should be stored at 4°C for up to 6 months.
Protocol
Cell Enrichment
This protocol provides a general guidance for binding to biotin-antibody and then enriching 105 cells using MagVigen™-Streptavidin magnetic nanoparticles. Please adjust the quantity of reagents for specific application.
- Gently vortex or pipette the MagVigen™- Streptavidin magnetic nanoparticles in the vial before use.
- Aliquot 20-50 μl nanoparticle solution for enrichment experiment. Magnetically 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 nanoparticles with 50 μl of Washing Buffer once. Magnetically 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. Re-disperse nanoparticles in 50 ul of Antibody Binding Buffer.
- Add 1µg-2.5µg biotin-conjugated antibody in a volume of 100μl to the nanoparticle and incubate for 30-60 minutes on a rotator.
- Wash nanoparticle-antibody conjugates with 100 μl of Wash Buffer twice to remove unbound antibody.
Resuspend the nanoparticle-antibody conjugates in 50 ul of Wash buffer, ready for downstream enrichment experiments. - Add the nanoparticles to the cell sample (100-200 ul), pipette mixing up and down for a few times. Then set the mixture on an orbital rotator for 30 minutes at room temperature.
- After incubation, use a magnet to separate the nanoparticles (with bound cells) from the solution, and carefully remove the supernatant.
- Wash the nanoparticle-cell complex with 200μl cell culture medium twice.
- Isolated cells can be re-suspended in cell culture medium or your choice of assay buffer for downstream applications.
Note: 20 μl nanoparticles could bind ~1µg of antibody.
Application for Cell Separation and Enrichment:
Note: 20-50 μl of nanoparticle sample is generally sufficient for the enrichment of 1-10×105 cells. Cell capture efficiency can be affected by factors such as distribution of target cells in the cell population, density of antigen/epitope expressed on the cell surface, and the antibody affinity. Optimize the input quantity of nanoparticle solution through an initial titration experiment is a good practice.
