Column and spin cup kits for purifying glutathione-S-transferase (GST) from bacteria.

B-PER GST Fusion Protein Purification Kits offer rapid purification of GST and GST-tagged fusion proteins from bacteria, yielding up to four times as much protein as other kits. The kits use B-PER Bacterial Protein Extraction Reagent to gently and efficiently lyse E. coli cells to extract soluble proteins. The B-PER Lysate is directly loaded on the pre-packed glutathione agarose column or incubated as a slurry in a spin cup. After washing to remove non-GST components of the lysate, the purified GST or GST fusion protein is eluted with reduced free glutathione.

Two kit formats provide the range of scale and convenience required for typical laboratory applications:

• The Column Kit purifies >10 mg of overexpressed GST protein per use in less than 3 hours.
• The Spin Kit purifies approximately 1 mg of overexpressed GST protein in 30 minutes.

B-PER GST Fusion Protein Column Purification Kit Highlights

• Capacity – purify >10 mg of overexpressed GST fusion protein from 250 ml bacterial culture; yields up to four times as much protein as competitor kits (Figure 1)
• Purity – yields GST fusion protein free of cellular contamination (Figure 2)
• Speed – progress from lysis to purified protein in less than two hours
• Easy and efficient cell lysis – no sonication required; complete cell lysis achieved with B-PER Bacterial Protein Extraction Reagent
• Ready-to-use columns and reagents – pre-packed columns (contain 1 ml glutathione agarose), prepared wash buffers and easy-to-prepare glutathione solution for elution make set-up and execution very easy
• Convenient size – five columns and sufficient reagents for five purifications

Figure 1. Comparison of the B-PER GST Fusion Protein Column Purification Kit vs. Brand P Kit. The pure protein yield from the B-PER GST Kit is approximately four times higher than that of Brand P. L = Lysate. W = Washes. E = Elution fractions. Note that most of the GST comes off in the first elution with the B-PER GST Kit, while the majority of the GST protein is found in the second elution fraction with Brand P’s kit.		Figure 2. SDS-PAGE analysis of GST purified using the Column Kit. Fractions from each purification step were analyzed by SDS-PAGE (4%-20%) and stained with GelCode Blue Stain. Lane 1: crude E. coli lysate extracted with B-PER Reagent; Lane 2: flow-through from crude lysate; Lanes 3-4: fractions of Wash Buffer 1; Lane 5: fractions of Wash Buffer 2; and Lane 6: GST eluted from the column with elution buffer (50 mM glutathione). Lane M: MW markers.

B-PER GST Fusion Protein Spin Purification Kit Highlights

• Speed – progress from E. coli cell culture to purified protein in less than 30 minutes!
• Purity – yields GST fusion protein free of cellular contamination (Figure 3)
• Capacity – purify > 1 mg of overexpressed GST fusion protein from 250 ml bacterial culture (Table 1)
• Easy and efficient cell lysis – no sonication required; complete cell lysis achieved with B-PER Bacterial Protein Extraction Reagent
• Ready-to-use components and reagents – microcentrifuge spin cups, glutathione agarose slurry, prepared wash buffer and easy-to-prepare glutathione solution for elution make set-up and execution very easy
• Convenient size – 16 spin cups and sufficient reagents for 16 purifications

Figure 3. Purification of GST using the B-PER GST Spin Purification Kit. Recombinant GST expressed in E. coli BL21 was first extracted with B-PER Reagent (Lane 1). After binding to the supplied immobilized glutathione (Lane 2: non-bound fraction), the GST-bound agarose beads were transferred to spin columns and washed once with wash buffer to remove contamination (Lane 3: recovered wash fraction). Finally, the recombinant protein was eluted four times to achieve complete elution (Lanes 4-5: elution fractions 2 and 3 of GST. Lane M: MW marker.

* B-PER Technology is protected by U.S. Patent # 6,174,704

Related Products
B-PER Bacterial Protein Extraction Reagents
Pierce Spin Cups and Columns
Glutathione Coated Microplates