BN QuickPick^TM GST
For the rapid fractionation of protein

QuickPick™ GST Magnetic Particles are formed from magnetic agarose particles which provide an extensive porous surface area especially suitable for the purification of recombinant GST-fusion proteins from cell lysates. The transfer of magnetic particles with PickPen® technology enables rapid purification of recombinant GST-fusion proteins. The methodology removes the need of using time-consuming column technology.

Glutathione-S-transferase (GST) is one of the most common affinity methods used to simplify the purification and detection of recombinant proteins expressed in Escherichia coli (E. coli). The principle of the mechanism is based on the interaction between a glutathione molecule covalently bound to magnetic particles via a spacer arm and a recombinant GST-fusion protein. An affinity elution using reduced glutathione liberates the GST-fusion proteins from the magnetic particles.

BN QuickPick^TM GST  protocol
Number tubes from 1 to 5 and pipette liquids before starting as follows:
Tube 1 :  400 µl GST Magnetic Particles
Tube 2 :  800 µl sample
Tube 3 :  800 µl GST Wash Buffer
Tube 4 :  800 µl GST Wash Buffer
Tube 5 :  100 µl GST Elution Buffer

Protocol

1.    Pick up the PickPen® tip using the PickPen®. Extend the magnet 2-3 times to check that the tip is firmly in place. Collect the magnetic particles from Tube 1 (Magnetic Particles) and release them into Tube 2 (sample). If the magnetic particles have formed clumps, they can be broken up by using the PickPen® tip. Mix briefly and gently. Note that the magnet has to be withdrawn at this point. Incubate for at least 5 minutes. Mix the solution occasionally to avoid sedimentation.

Incubate for at least 5 minutes

2.      Collect the magnetic particles from Tube 2 and wash them in Tube 3 (Wash Buffer) by releasing the magnetic particles into the solution and mixing gently.  Repeat the washing in Tube 4. Collect the particles from Tube 4 (start collecting the magnetic particles from the very surface of the liquid to ensure that they collect only onto the end of tip) and release carefully into Tube 5 (Elution Buffer). Rub the tip against the tube wall to ensure that all magnetic particles are released into the suspension.  Mix the magnetic particles thoroughly but gently in the solution and incubate for at least 5 minutes. Mix the solution occasionally to avoid sedimentation.

Incubate for at least 5 minutes

3.    After incubation collect the magnetic particles from Tube 5 and discard. The purified GST-tagged protein is ready to be used for downstream applications.

For details of the PickPen^TM magnetic transfer device click here

FOR RESEARCH USE ONLY - NOT FOR USE IN DIAGNOSTIC PROCEDURES

Metachem Diagnostics Ltd.
29 Forest Road, Piddington, Northampton. NN7 2DA
Tel. (01604) 870370     Fax (01604) 870194