Search
Search
View additional product information for Dynabeads™ MyOne™ Streptavidin T1, for OEM and industrial use only - FAQs (35601)
16 product FAQs found
We do not recommend this as streptavidin becomes hydrophobic and aggregates during denaturation.
Which product to choose depends on the properties of your sample, the buffers and solutions applied, as well as the downstream application. In general, all Dynabeads Streptavidin beads can be used in applications involving biotinylated ligands; however, some beads may perform better than others in particular applications due to their characteristics. Dynabeads M-280 Streptavidin beads and Dynabeads MyOne Streptavidin T1 beads are commonly used for protein and nucleic acid applications. Dynabeads M-270 Streptavidin beads and MyOne Streptavidin C1 beads are preferred for nucleic acid diagnostics, specifically with samples that have a high concentration of chaotropic salts, immunoassays involving small biotinylated antigens, and in applications that are not compatible with BSA, as these beads are not blocked with BSA. Dynabeads MyOne Streptavidin beads offer increased binding capacity and slower sedimentation rate, making them ideal for automated applications and when larger amounts of a biotinylated compound or its specific target need to be isolated. Please see the selection guide here ( https://www.thermofisher.com/us/en/home/brands/product-brand/dynal/streptavidin-coupled-dynabeads.html?icid=fr-strep-1).
You can assay the supernatant for unbound nucleic acid to determine the amount of nucleic acid bound to the Dynabeads Streptavidin beads. The concentration of nucleic acids can be checked by measuring the OD or by running them on a gel. Alternatively, the nucleic acids can be labeled radioactively and the concentration measured directly on the beads, or fluorescently and measured in the supernatant.
Our Dynabeads Streptavidin magnetic beads can be used directly in PCR or real-time PCR. However, you would have to empirically optimize the amount of beads to be used per volume of reaction.
The exact number of streptavidin molecules bound per bead is not measured, but is approximately 14-16 µg streptavidin per milligram Dynabeads M-280 Streptavidin magnetic beads.
All biotin reagents should contain a spacer arm, at least a 6-carbon linker, to reduce steric hindrance. This is because the bicyclic ring of biotin goes deep into the biotin binding cleft in streptavidin. A 6-carbon arm is the minimum length between biotin and the first base of sequence that is required to reduce the steric hindrance effect. The longer this distance is, the less the steric hindrance. A 6-carbon linker is standard linker size from most companies and should be enough for most applications. We recommend specific biotinylation at the 5'-end of the probe.
In direct capture, the biotinylated probe/ligand is first coupled to the Dynabeads magnetic beads followed by addition of your sample. In indirect capture, the biotinylated probe/ligand is first added to the sample followed by addition of your Dynabeads magnetic beads. Precoupled ligand for direct capture allows you to reuse the Dynabeads magnetic beads, while an indirect approach can be beneficial when the concentration of your target is low, specific affinity is weak, or the binding kinetics is slow. Please see the following link (https://www.thermofisher.com/us/en/home/life-science/dna-rna-purification-analysis/napamisc/capture-of-biotinylated-targets/nucleic-acid-capture-assays.html) for a diagram of the capture.
- One milligram of Dynabeads M-280 Streptavidin magnetic beads typically binds 650-900 pmol of free biotin, 200 pmol of biotinylated peptide, up to 10 µg of biotinylated antibody, 10 µg of biotinylated double-stranded DNA, or 200 pmol of biotinylated single-stranded oligonucleotides.
- One milligram of Dynabeads M-270 Streptavidin magnetic beads typically binds more than 950 pmol of free biotin, 200 pmol of biotinylated peptide, up to 10 µg of biotinylated antibody, 10 µg of biotinylated double- stranded DNA, or 200 pmol of biotinylated single-stranded oligonucleotides.
- One milligram of Dynabeads MyOne Streptavidin C1 magnetic beads typically binds more than 2,500 pmol free biotin, 400 pmol of biotinylated peptides, up to 20 µg of biotinylated antibody, 20 µg of biotinylated double-stranded DNA, or 500 pmol of biotinylated single-stranded oligonucleotides.
-One milligram of Dynabeads MyOne Streptavidin T1 magnetic beads typically binds 1,100-1,700 pmol free biotin, 400 pmol of biotinylated peptides, up to 20 µg of biotinylated antibody, 20 µg of biotinylated double- stranded DNA, or 400 pmol of biotinylated single-stranded oligonucleotides.
Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.
Storage should be at 2 to 8 degrees C. Freezing or drying of the Dynabeads magnetic beads is not recommended. Provided the Dynabeads magnetic beads are stored correctly, quality is guaranteed until the expiry date stated on the label.
Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.
The binding capacity of streptavidin-coupled Dynabeads magnetic beads is fragment length-dependent. Reduced binding capacity for large DNA fragments may be due to steric hindrance. For large DNA fragments (greater than 2 kb in size), we recommend using Dynabeads kilobaseBINDER Kit.
-Salt concentration affects the binding efficiency of biotinylated nucleic acids to the Streptavidin-coupled Dynabeads magnetic beads. Optimal binding conditions for biotinylated DNA fragments (up to 1 kb) are achieved at 1 M NaCl (final concentration), 25 degrees C and 15 min incubation time. Longer DNA fragments should be immobilized overnight. Biotinylated antibodies should be immobilized in PBS buffer pH 7.4, supplemented with 0.1% BSA.
-Ensure that your sample does not contain an excess of free biotin, as the free biotin will bind Streptavidin-coupled Dynabeads magnetic beads much more rapidly than larger biotinylated molecules. Biotinylated oligonucleotides should be recovered by reverse phase HPLC or FPLC to remove free biotin from the sample.
-We also recommend a titration to optimize the quantity of beads used for each individual application, since both the size of the specific molecule to be immobilized and the biotinylation procedures affect the binding capacity of the beads.
Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.
Use mild elution conditions, e.g., a buffer with high salt or low pH. Heating the beads at 95 degrees C for 5 mins in SDS sample buffer will elute the antibody as well.
Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.
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. Biotin-streptavidin interactions are more easily reversible when biotin derivatives with lower binding affinity are used along with chemically modified streptavidins with lower biotin binding affinity. When modified biotins and streptavidins are used, gentle methods for inducing reversible binding are available
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.
Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.
We do not recommend this because streptavidin becomes hydrophobic and aggregates during heat-induced denaturation.
Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.
The product to choose depends on the properties of your sample, the buffers and solutions you will use, as well as your downstream application. In general, all the Dynabeads Streptavidin beads can be used in applications involving biotinylated ligands. However, some beads may perform better than the others in particular applications due to their characteristics. Please refer to the Dynabeads Streptavidin Selection Guide (https://www.thermofisher.com/us/en/home/brands/product-brand/dynal/streptavidin-coupled-dynabeads.html#selection) for detailed information.
Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.
The streptavidin molecule is covalently attached to the surface of the beads; under normal, recommended conditions, negligible leakage is detected (less than 0.2% of total attached streptavidin after 2 months at 37 degrees C). However, it should be noted that not all of the four streptavidin subunits are covalently coupled to the beads. Typically, one or two of the subunits are covalently coupled. Streptavidin is like other proteins; if heated, it can denature and dissociate into subunits. If Dynabeads Streptavidin magnetic beads are, for instance, boiled, some of the streptavidin subunits may be released (as monomers or aggregates) from the beads. The covalently bound streptavidin subunits will not be affected by such treatment. When streptavidin is bound to biotin, the streptavidin-biotin complex is more stable than the unbound streptavidin molecule
Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.
The exact number of streptavidin molecules bound per bead is not measured, but is approximately 14-16 µg streptavidin per mg Dynabeads M-280 Streptavidin magnetic beads.
Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.