Glutathione Agarose Beads
Glutathione Agarose Beads

Glutathione S-transferase (GST) gene fusion systems have been widely used for obtaining large amounts of desirable protein in Escherichia coli.
 
Name Cat# Description Price Qty
Glutathione Agarose
GAB-100
10 ml (20ml total volume with 50% suspension)
$94.04
Glutathione Agarose
GAB-200
25 ml (50ml total volume with 50% suspension)
$169.95
Glutathione Agarose
GAB-300
100 ml (200ml total volume with 50% suspension)
$509.85
Glutathione Agarose
GAB-500
500 ml (1000ml total volume with 50% suspension)
$2035.55
Glutathione Agarose
GAB-OEM
Any Size
please inquire

Description:
Glutathione S-transferase (GST) gene fusion systems have been widely used for obtaining large amounts of desirable protein in Escherichia coli. The fusion protein, which contains a GST tail can then be purified through affinity chromatography. MCLAB’s glutathione agarose is designed for the specific purification of GST recombinant proteins and other glutathione-binding proteins. Glutathione agarose is uniquely formulated for excellent binding capacity and purity of the protein of interest.


Specifications

Chromatography technique GST-tagged protein purification
Matrix Highly cross-linked 6% beaded agarose
Active group Glutathione
Active group density >40 umol/ml drained medium
Binding Capacity > 25 mg horse liver GST/ml resin
Spacer 1,4-bis(2,3-epoxypropoxy)butane (12 atom stable and uncharged ether hydrophilic linkage)
Bead geometry & size Spherical 50 to 150 um
Bead mean diameter d50v 90 um
pH Stability working range 4 to 13
pH Stability cleaning in place (cip) 4 to 13
Chemical stability Stable to commonly used aqueous solutions. Can be used with non-ionic detergents, denaturing solvents, e.g. 8 M urea and 6 M guanidine hydrochloride, Stable in organic solvents, e.g. 50% methylformamide and 50% dioxane.
Storage 2 to 8 degC in 20% Ethanol


Protocol:
The following instruction for GST-fusion protein purification can be scaled up or down depending on the user’s preference. This manual exemplifies sample preparation from a specific amount of starting material and purification using 1 ml resin.
1. Centrifuge sample after cell lysis to remove undissolved membranes and cellular debris before applying to the purification column.
2. Wash the purification column with 10x bead volume of Binding Buffer to remove azide.
3. Dilute an appropriate amount sample with a 1:1 ratio of Binding Buffer before applying to the purification column.
4. Wash the purification column with 10x bead volume of Binding Buffer or until no proteins can be detected in the washes.
5. Elute the bound protein of interest with 5x bead volume of Elution Buffer.
6. GST agarose beads can be saved for later use by washing the purification column with Binding Buffer containing 3 M NaCl. After a thorough wash, the purification column should be equilibrated in Binding Buffer containing 2 mM sodium azide and stored at 4°C.
GST-fusion proteins are generally eluted from glutathione agarose beads with the use of excess glutathione. Alternatively, GST-fusion protein can be encoded with a cleavage site between the GST and the protein, allowing the desirable protein to be eluted with the use of a protease.

Recommended Storage Conditions: 2ºC to 8ºC.

Binding Buffer:
50 mM Tris
150 mM NaCl
pH 8.0

Elution Buffer:
50 mM Tris pH 7.8
0.15 M NaCl
1 mM EDTA
1 mM DTT
10 mM GST

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