BN QuickPickTM IMAC
For the rapid fractionation of protein

    

QuickPickTM  IMAC (Immobilised Metal Ion Affinity Chromatography) magnetic particles are formed from magnetic agarose particles which provide an extensive porous surface area especially suitable for the purification of His-tag proteins from cell lysates.  The transfer of magnetic particles with the PickPenTM technology enables rapid purification of His-tag recombinant proteins with proper binding and elution characteristics.   The methodology removes the need of sample pre-treatment or use of time consuming column technology. 

Immobilised metal ion affinity chromatography (IMAC) was originally introduced as a method for group separation, but today it represents one of the most important tools for the single-step purification of proteins based on polyhistidine tags.  The principle of the mechanism is based on the interaction between a metal ion coordinated to a covalently bound chelating ligand and a His-tag protein.

BN QuickPickTM IMAC  protocol
Number tubes 1 to 6 
Tube 1 : 100 mL IMAC magnetic particles in storage solution
Tube 2 :  400 mL IMAC Regeneration Buffer
Tube 3 :  400 mL IMAC Wash Buffer 1
Tube 4 :  300 mL sample
Tube 5 : 400 mL IMAC Wash Buffer 2
Tube 6 :   50 mL IMAC Elution Buffer

Protocol
1.    Collect IMAC magnetic particles from Tube 1 with PickPen and regenerate them by releasing into tube  Tube 2 (IMAC Regeneration Buffer).
2.      Collect magnetic particles from Tube 2 with PickPen and release them into Tube 3 (IMAC Wash Buffer 1).  Vortex in pulses.
3.    Collect magnetic particles from Tube 3 with PickPen and release them into Tube 4  (sample).  Mix the magnetic particles thoroughly and incubate for 2 minutes.  Mix occasionally to avoid sedimentation.

4.    Collect the magnetic particles from Tube 4 with PickPen and wash them in Tube 5 (IMAC Wash Buffer 2) by releasing the particles into the solution.  Mix the suspension in Tube 5 for 10 seconds.       
5.    Collect the magnetic particles from Tube 5 with Pickpen and release them into Tube 6 (IMAC Elution Buffer) and mix thoroughly.  Incubate for at least 1 minute mixing  occasionally.
 

6.    After incubation collect the magnetic particles  from Tube 6 and discard.  The sample is now in the eluate and ready to be used for downstream applications.

 

Purification of His-tagged proteins
from bacterial cells.

 

QuickPickTM IMAC Catalogue Number   No of Preparations
IMAC Kit BN 62301 8 preps
IMAC Kit BN 62311 48 preps
IMAC Plus Kit* BN 62303 8 preps

*IMAC Plus purifies between 10 and 15 times more protein than the standard kit.

For details of the PickPenTM 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