Glutathione Agarose

October 02, 2017

University of South Alabama
Cancer Biology
Graduate Student

Overall

Quality of Results

Ease-of-Optimization

What do these ratings mean?
Write a Review

Company:

ThermoFisher Scientific

Product Name:

Pierce Glutathione Agarose

Catalog Number:

16100

The glutathione agarose beads take advantage of the high affinity interaction between glutathione and GST to pull-down GST fusion proteins from a mixture such as mammalian or bacterial cell lysates. I used the Pierce Glutathione Agarose to pull-down recombinant GST-RAF-RBD from E. coli lysates.

Experimental Design and Results Summary

Application

Affinity Purification

Starting Material

Mammalian or bacterial cell lysates

Protocol Overview

Wash the glutathione agarose beads twice with the same buffer as your sample. This is done by suspending the beads in the buffer, centrifuging the beads for 30 seconds at 6000 x g, and then removing the supernatant. Incubate your sample on a rotator with the beads for 1 hour at 4 degrees Celsius. Centrifuge the beads for 30 seconds at 6000 x g, and remove the supernatant. Then wash the beads three times as described previously. Finally, resuspend the beads in the buffer of your choice to achieve your desired volume.

Tips

Be sure to incubate the beads long enough with your sample to bind all of the GST fusion protein in your sample. Optimization of incubation time may be required.

Results Summary

The glutathione agarose beads allowed me to purify over a milligram of my GST fusion protein. The washing allowed me to remove unbound protein to have a product with high purity.

DOI or PMID #

N/A

Additional Notes

N/A

Image Gallery

Summary

The Good

Much easier to use for purifying protein than other methods.

The Bad

The beads are very viscous, so be careful that your pipet tip is not clogging in order to ensure you're pipetting proper volumes.

The Bottom Line

The glutathione agarose beads from ThermoFisher are of high quality and make purifying protein very simple. The same to be very stable even when freezing at -80 degrees Celsius. They are also affordable compared to other purification methods.

Join the discussion