DUALrefold system

The DUALrefold system: efficient refolding of denatured cytosolic proteins and integral membrane proteins

Refolding denatured proteins is often difficult

Often, proteins can only be purified from total cellular extracts using denaturing conditions. But even when applying mild conditions, some proteins can only be purified in a denatured state. This is particularly true for larger proteins with complicated structures, such as membrane proteins.

The principle of protein refolding is based on the following parameters:

Different proteins require different conditions to fold
Conditions that stabilize the folding intermediates often help to yield correctly folded proteins
Columns completely remove denaturants

The DUALrefold system integrates all these three factors and provides a refolding kit which is both very efficient and easy to use.

DUALrefold kits and reagents help to refold proteins efficiently

The DUALrefold kits help you to select the right conditions to refold your purified protein to an active state. Initially, a large number of different refolding conditions are screened using the spin columns supplied in the DUALrefold kit. Once the optimal refolding conditions have been identified, the appropriate refolding column can be used for large scale preparation of refolded protein.

Refolding of cytosolic (soluble) proteins

DUALrefold soluble protein refolding kit

Using the DUALrefold soluble protein refolding kit, optimal protein folding conditions can be identified by a simple bench-top centrifugation and incubation procedure. No tedious dialysis or high ratio dilution is needed. No special equipment or protein folding experience is required to use the folding kit. About 0.06 - 0.12 mg of inactive protein can be used per refolding assay.

Small scale refolding columns

Small scale DUALrefold protein folding columns are designed for small-scale (0.6 - 1.2 mg) preparative protein folding experiments after the optimal folding conditions have been identified. The column set includes reagents and 10 identical protein folding spin-columns.

Large scale refolding columns

Large scale DUALrefold protein folding columns are designed for preparative protein folding experiments (5 -10 mg protein) after the optimal folding conditions have been identified. The column set includes 4 identical preparative protein folding columns and reagents.

Refolding of integral membrane proteins

DUALrefold membrane protein refolding kit

The DUALrefold membrane protein refolding kit is specifically designed for refolding of membrane proteins from urea-solubilized inclusion bodies or other protein aggregates. Using the supplied spin columns and reagents, 20 different refolding conditions (including various detergents and lipids that form micelles and bicelles) can be tested. The micellar and bicellar environments facilitate folding receptors, ion channels and other membrane proteins. Preparative columns are available once optimal refolding conditions have been identified.

DUALrefold membrane protein refolding columns

The DUALrefold membrane protein refolding column sets are used for preparative membrane protein folding under optimized folding conditions. Each column set includes 4 identical preparative protein folding columns and reagents for folding about 5 - 10 mg of urea-solubilized membrane proteins.