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Isolation and analysis of GST-tagged proteins
Background
The glutathione S-transferase (GST) enzyme family comprises a long list of cytosolic, mitochondrial, and microsomal proteins which are capable of multiple reactions with a multitude of substrates, e.g. glutathione. Genetic engineers have used glutathione S-transferase to create the so-called GST gene fusion system. Here, the GST tag is used to purify and detect glutathione-binding proteins of interest.

A high-quality monoclonal Anti-GST antibody coupled to µMACS™ MicroBeads is offered by Miltenyi Biotec, allowing specific isolation of recombinant GST fusion proteins on an analytical scale.
For medium- to large-scale purification of GST-tagged proteins, the ProCatch Glutathione Resin is recommended (fig. 1).
Isolation and purification of GST-tagged proteins
The µMACS™ and MultiMACS™ GST Isolation Kit take advantage of the well-established MACS® Technology; thus, allows fast and specific isolation of pure GST-tagged proteins. The advantages of MACS Technology for protein isolation are:

Specificity – high-quality monoclonal antibodies coupled to MACS MicroBeads assure specific isolation of GST-tagged proteins with low background.
Sensitivity – fast reaction kinetics of tiny (50 nm), non-sedimenting MACS MicroBeads allow sensitive isolation even when working with rare proteins.
Speed – less than 2 hours to high-purity GST-tagged proteins!

The µMACS GST Isolation Kit was developed for manual, low-throughput applications with the µMACS Separator.
The procedure can easily be upscaled with the MultiMACS GST Isolation Kits to a semi- or fully-automated, high-throughput processing of up to 96 samples in parallel by utilizing the MultiMACS 96 Separator.

For medium- to large-scale purification of GST-tagged proteins the ProCatch Glutathione Resin is recommended.

Antibody specifications
Target sequence:
whole glutathione S-transferase (220-residue polypeptide)
Tag origin: Schistosoma japonicum
Isotype: mouse IgG1
Columns
For µMACS™ Isolation Kit: µ Column, upscale with M Column

For MultiMACS™ Isolation Kits: Multi-8 or Multi-96 Columns
Further information
Protein isolation flyer.pdf
[PDF; 814,4 KB]
 
Figure 1
Purification of a 30 kDa glutathione-binding protein with ProCatch Glutathione Resin.
A Marker (lane 1), lysate from cell extract (2), flow-through (3), first eluate (4), and second eluate (5) were separated by SDS-PAGE and Coomassie stained. B Low-pressure column purification protocol. Peak 1 defines non-bound protein fraction (flow-through). Peak 2 corresponds to the eluted glutathione-binding protein. Absorbance was measured at 280 nm.
A
B
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Products
μMACS GST Isolation Kit
Tag specification
- for 40 isolations
Components
- 2 mL μMACS GST MicroBeads
- 2×50 mL Lysis Buffer
- 50 mL Wash Buffer 1
- 5 mL Wash Buffer 2
- 5 mL Elution Buffer
Download datasheet
130-091-370
Qty.:
 

μMACS Anti-GST Starting Kit
- for 40 isolations
Components
- 1 μMACS GST Isolation Kit
- 1 μMACS Separation Unit
- 1 MACS Multistand
- 2×20 Columns
130-091-493
Qty.:
 

MultiMACS GST Isolation Kit (12×8)
- for 96 isolations
Components
- 3x2 mL µMACS Anti-GST MicroBeads
- 1x50 mL Equilibration Buffer
- 1x Multi-8 Column Box (12x8)
Download datasheet
130-094-254
Qty.:
 

MultiMACS GST Isolation Kit (4×96)
- for 384 isolations
Components
- 5x4.6 mL µMACS Anti-GFP MicroBeads
- 1x50 mL Equilibration Buffer
- 1x Multi-96 Column Box (4x96)
Download datasheet
130-094-256
Qty.:
 

ProCatch Glutathione Resin
- 10 mL
Download datasheet
130-092-187
Qty.:
 

- 25 mL
Download datasheet
130-092-186
Qty.:
 

- 100 mL
Download datasheet
130-092-185
Qty.:
 

Related products
Isolation and analysis of c-myc-tagged proteins
Isolation and analysis of GFP-tagged proteins
Isolation and analysis of HA-tagged proteins
Isolation and analysis of His-tagged proteins
μMACS™ and MultiMACS™ Protein A or Protein G Kits
μMACS™ and MultiMACS™ Streptavidin Kits
References
1. Paz et al. (2005) Mol Cancer Ther 4(11): 1801-1809.
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