Efficient, high yield chromatography purification of biotin and biotinylated substances. Binding capacity: > 300 nmol (75 μg) biotin / ml of settled resin. ... Read more
The ProteIndexTM Streptavidin Agarose 6FF provides a simple, rapid, and reliable method to purify the biotin and biotinylated substances. The streptavidin ligand is a recombinant version of the native protein that is originated from Streptomyces avidinii, and has an extraordinarily high affinity for biotin. The immobilized streptavidin on the highly cross-linked 6% agarose matrix support can therefore be used for affinity chromatography purification of biotin and biotinylated substances. Due to strong interaction between streptavidin and biotin, elution of biotin and the biotinylated substance usually requires harsh denaturing conditions. Streptavidin also binds to 2-iminobiotin and iminobiotinylated substances, in a pH-dependent fashion. The affinity interaction between the streptavidin and iminobiotin is strong when the pH is above 9.0 and decreases dramatically as the pH is lowered. Therefore, the iminobiotinylated substance can be captured onto the immobilized streptavidin at pH 9.5 – 11.0 and can be eluted at pH 4.0.
Alternatively, the immobilized streptavidin on the matrix support can be used to purify biomolecules that are able to interact and form complexes with a biotinalyted biomolecules. For example, an antigen can be purified via the immunoreactive complex with its specific, biotinylated antibody captured onto the Streptavidin Agarose 6FF resin, and the antigen can be eluted when it dissociates from the complex.
The base matrix of Streptavidin Agarose 6FF is highly cross-linked 6% agarose, providing excellent flow properties, and high mechanical stability. The resin is well suited for batch/gravity purification, centrifugation procedure, or FPLC process on automated systems.
- High selectivity for high purity
- High capacity: binding > 75 μg (300 nmol) Biotin / ml of settled resin
- Suited for batch, gravity, spin, and automated systems.
Figure 1. Purification performance of Streptavidin 6FF resins.
E.coli lysate containing an iminobiotinylated protein (~ 45 kDa) was 1: 5 diluted in Binding Buffer (50 mM (NH4)2CO3, 0.5 M NaCl, pH10.0) and then was loaded on 0.5 mL packed Streptavidin 6FF resins. After washes with the Binding Buffer, bound material was eluted in Elution Buffer (50 mM NH4Ac, 0.5 M NaCl, pH 4.0 ). The resulted resin was then incubated with SDS sample buffer at 95°C for 10 minutes. Fractions were analyzed by SDS-PAGE.
Lanes from left to right: Molecular Weight markers (MW), Cell lysate (CL), Flow-through (FL), and Eluate (E), and the Supernatant of boiled resin in SDS sample buffer (B).
|Ligand||Recombinant Streptavidin expressed in E.coli
> 75 μg (300 nmol) Biotin / ml of settled resin
> 6 mg Biotinylated serum albumin / ml of settled resin
|Matrix||6% highly cross-linked agarose supplied as 50% slurry|
|Particle size||45 μm - 165 μm|
||0.3 MPa (3 bar, 43 psi)|
|Recommended flow rate
||300 - 500 cm/hour|
||pH3 - 10 (short term) and pH4 - 9 (long term)
||2 - 8°C|
||20% ethanol as preservative|
Product is shipped at ambient temperature. Upon receipt store at 2 - 8°C. Do not Freeze.
Marvelgent Biosciences Inc.
116 Will Drive, Canton, MA 02021, USA
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