What is Glutathione Agarose?

Thermo Scientific Pierce Glutathione Agarose is a high-capacity, high-performance resin for affinity purification of GST-tagged fusion proteins from cellular lysates. Features of Glutathione Agarose: • Slurry—crosslinked 6% beaded agarose, slurried in water with sodium azide.

How do you regenerate GST beads?

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  1. After the elution step, add 10 bv Wash Buffer and allow the volume to completely flow through the matrix.
  2. Rinse the column again with 10 bv water.
  3. Wash with 2 bv of one of the following solutions.
  4. Rinse the column again with 10 bv water.
  5. Wash the column with 10 bv Wash Buffer.

How do you elute GST fusion protein?

GST-fusion protein elution-can anyone help?

  1. Centrifuge 9000 rpm/500 g for 5 min at RT.
  2. Wash beads three times (9000 rpm/500 g for 5 min at RT) with 1X PBS containing 1% Triton X-100.
  3. Add 10/20 mM reduced glutathione.
  4. Centrifuge at 9000 rpm/500 g for 5 min to sediment the gel, and remove the supernatant.

What is glutathione resin?

Glutathione Resin (Cat. No. L00206) is an affinity chromatography medium designed for easy, one-step purification of recombinant glutathione S-transferase (GST) fusion proteins and other glutathione binding proteins expressed in E. coli, insect cells and mammalian cells.

What is a GST pull down assay?

GST pull-down assays involve affinity purifications of one or several unknown proteins from a biological sample using a GST-tagged bait protein. The basic principle is that the GST-tagged bait protein binds to its partners, and the resulting complex is captured on beads with immobilized glutathione.

How do you make 10mm glutathione?

  1. Centrifuge 9000 rpm/500 g for 5 min at RT.
  2. Wash beads three times (9000 rpm/500 g for 5 min at RT) with 1X PBS containing 1% Triton X-100.
  3. Add 10/20 mM reduced glutathione.
  4. Centrifuge at 9000 rpm/500 g for 5 min to sediment the gel, and remove the supernatant.
  5. Repeat elution and centrifugation steps twice more.

How do you clean glutathione beads?

Wash the Glutathione Sepharose 4B pellet with 10 bed volumes* of 1X PBS. Centrifuge the suspension at 500 x g for 5 minutes to sediment the gel. Discard the wash. Repeat the wash twice for a total of three washes.

What are glutathione levels?

Glutathione is an antioxidant produced in cells. It’s comprised largely of three amino acids: glutamine, glycine, and cysteine. Glutathione levels in the body may be reduced by a number of factors, including poor nutrition, environmental toxins, and stress. Its levels also decline with age.

What is the difference between IP and co IP?

In immunoprecipitation (IP), an antibody is used to purify its specific target, or antigen from a mixture. In co-immunoprecipitation (Co-IP), an antibody is used to purify its target antigen, along with its binding partners, from a mixed sample.