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Description
Thermo Scientific Pierce Streptavidin Magnetic Beads accelerate throughput for automated magnetic purification of biotinylated molecules.
Features of Streptavidin Magnetic Beads:
- High-performance beads—non-aggregating, pre-blocked, iron oxide, superparamagnetic microparticles provide exceptional uniformity for automated HTS and manual applications alike
- Stable immobilization chemistry—streptavidin is immobilized using leach-resistant chemistry
- High capacity—superior quality beads with high binding capacity provide rapid and efficient biomolecule purification from complex samples
- Low non-specific binding—stable, pre-blocked beads provide clean purification products (e.g., antigen eluted in IP with biotinylated antibody) that are compatible with mass spectrometry analysis and other downstream analyses
- Superior performance—nearly three times higher binding capacity than typical beads from other suppliers, allowing the use of smaller amounts per experiment
Applications:
- Immunoprecipitate antigens (using biotinylated antibodies) from a wide variety of sources
- Co-immunoprecipitate interaction complexes using biotinylated antibodies
- Capture protein-protein interactions in pull-down assays using biotinylated 'bait' proteins
- Isolate biotin-labeled DNA-protein complexes from cell or tissue extracts
- Capture single-stranded biotinylated DNA oligos
- Isolate biotinylated PCR products
These streptavidin magnetic beads are validated and optimized for use with high-throughput magnetic platforms, such as the Thermo Scientific KingFisher 96 and KingFisher Flex Instruments, but the beads also enable premium performance for simple benchtop applications using an appropriate magnetic stand. The iron oxide, super-paramagnetic particles offer superior performance (high capacity and low nonspecific binding) compared with other commercial magnetic beads.
Pierce Streptavidin Magnetic Beads use a recombinant form of streptavidin with a mass of 53kDa and a near-neutral isoelectric point (pI). The protein is a tetramer having four biotin-binding sites. Unlike avidin, streptavidin has no carbohydrate groups, resulting in low nonspecific binding. The high-affinity interaction between streptavidin and biotin cannot be dissociated efficiently except with very harsh conditions, such as boiling in sample loading buffer for SDS-PAGE or 8M guanidine·HCl, pH 1.5. Consequently, it is often possible to elute binding partners in an interaction complex without also eluting the biotinylated component.
Specifications
Specifications
| Capacity (Metric) | 1 mL |
| Concentration | Slurry: 10 mg/mL, 1% solids |
| Content And Storage | Upon receipt store at 4°C. Product is shipped on an ice pack. |
| Particle Size | 1 μm |
| Form | Liquid |
| Product Line | Pierce |
| Quantity | 1 mL |
| Protein Form | Recombinant |
| Target | Biotin |
| Type | Magnetic Affinity Bead |
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Frequently Asked Questions (FAQs)
The streptavidin-biotin interaction is the strongest known non-covalent biological interaction between a protein and a ligand. Binding of biotin and streptavidin is very rapid and, once formed, the complex is unaffected by wide extremes of pH, temperature, organic solvents, and other denaturing agents. Normally, very harsh methods are required to dissociate the biotin from streptavidin, which will be irreversibly denatured by the procedure.
To dissociate biotinylated proteins from streptavidin, boil the beads in 0.1% SDS or SDS-PAGE buffer for 3 mins or incubate them in 8 M guanidinium hydrochloride, pH 1.5.
There are two methods to dissociate the non-biotinylated DNA from the biotinylated DNA strand. The following protocols are based on using 20 µL of Dynabeads Streptavidin, but are scalable. Both methods may release very small amounts of complementary biotinylated strand from streptavidin. If it is critical that no biotinylated strand is released, either adopt a different biotin modification using dual biotin (two biotin groups in sequence) or covalently bind DNA to e.g., Dynabeads M-270 Carboxylic Acid.
Using heat:
- Wash the DNA coated Dynabeads in 50 µL 1 x SSC.*
- Resuspend the beads in another 50 µL of 1 x SSC Incubate at 95 degrees C for 5 mins.
- Quickly put the tube in magnet stand for 1-2 mins and transfer the supernatant to a new tube.
- The supernatant contains your non-biotinylated DNA strand.
Using NaOH:
- Wash the DNA coated Dynabeads in 50 µL 1 x SSC.*
- Resuspend the beads in 20 µl of freshly prepared 0.15 M NaOH.
- Incubate at room temperature for 10 mins. Put the tube in magnet stand for 1-2 mins and transfer the supernatant to a new tube.
- The supernatant contains your non-biotinylated DNA strand. Neutralize the probe by adding 2.2 µL 10 x TE, pH 7.5 and 1.3 µL 1.25 M acetic acid.
Wash the Dynabeads coated with biotinylated strand once with 50 µL 0.1M NaOH, once with 50 µL of B&W buffer and once with 50 µL TE buffer.
*1 x SSC (0.15 M NaCl, 0.015 M sodium citrate. Dissolve the reagents in 800 mL water. Adjust pH to 7.0 with NaOH. Adjust the volume to 1 liter with water).
Assay the supernatant for unbound molecules. This will determine the amount of molecule bound to the Dynabeads. For nucleic acids, the concentration can be checked by OD readings, or by running a gel. For proteins, the concentration in the supernatant can be determined by a spectrometer using a protein assay like BCA. Alternatively, you can label the molecule with radioactivity or fluorescence and measure the concentration of molecule directly on the beads (former) or in the supernatant (latter).
Streptavidin is a protein composed of four identical subunits, each containing a high affinity binding site for biotin (K-D = 10 -15 M) . Streptavidin has the same biotin binding properties as avidin, but it has a low isoelectric point (pI=5) and no carbohydrate groups, resulting in low non-specific binding.
The streptavidin in Pierce Streptavidin Magnetic Beads is not His-tagged.
For Research Use Only. Not for use in diagnostic procedures.
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