Efficient, single-step chromatography purification of Maltose binding protein (MBP) tagged fusion proteins with high yield and purity. Stable and resistant to amylase degradation. ... Read more
PROTEINDEX™ Dextrin Agarose 6FF is a chromatography medium for purifying proteins fused to maltose binding protein (MBP-tagged protein). Recombinant proteins with the MBP-tag often have increased expression levels and higher solubility. Proper folding of the attached protein has also been shown to be promoted by the MBP tag. Since MBP increase the solubility, the tag is extremely useful for recombinant proteins that are prone to be accumulated in an insoluble form (inclusion bodies).
Dextrin is the polymeric product resulted by enzymatic hydrolysis of amylose with beta amylase. Compared to Amylose that is susceptible to amylase degradation, Dextrin is very stable. When used as the ligand substrate immobilized on the resin for purification of the MBP tag, Dextrin offers consistently strong affinity absorption even after repeated uses.
Affinity purification using Dextrin Agarose 6 Fast Flow take place under physiological conditions and mild elution is performed using maltose which preserve target protein activity. These mild conditions may even allow purification of intact protein complexes.
The base matrix of Dextrin Agarose 6 Fast Flow is a robust, highly cross- linked 6% agarose. The crosslinking of the base matrix has been optimized to give high physical and chemical stability, high specificity of the binding.
Our proprietary ProteIndex™ coupling technology ensures stable immobilization of active ligand and minimal ligand leakage in a wide range of purification buffer conditions. The resins can be reused multiple times to purify the same target protein.
- High selectivity for high purity
- > 10 mg MBP tagged protein (80 kDa)/mL medium
- Stable coupling of ligand, low leakage; can be re-used to purify the same protein
Figure 1. Purification performance of Dextrin 6FF resins.
50 mL of E.coli lysate containing a MBP-tagged fusion protein (~ 80 kDa) was loaded on 0.5 mL packed Dextrin 6FF resins at 0.5 mL/mL. The column was then washed with 20 mL of Binding Buffer (20 mM Tris-HCl, 200 mM NaCl, 1 mM EDTA, pH7.4). The target MBP-tagged protein was eluted with 9.5 mL of elution buffer (20 mM Tris-HCl, 200 mM NaCl, 1 mM EDTA, 10 mM maltose, pH7.4). Fractions of lysate, flow-through, and eluate were analysed by SDS-PAGE. Lanes: (1) Raw cell lysate; (2) Flow-through; (3) Eluate.
|Binding capacity||>10 mg MBP tagged protein (80 kDa)/mL medium|
|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|
||2 - 8°C|
||20% ethanol as preservative|
Product is shipped at ambient temperature. Upon receipt store at 2 - 8°C. Do not Freeze.
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