Glutathione Magnetic Beads for GST Pull-Down Assay

Product

Glutathione Magnetic Beads for GST Pull-Down Assay

Catalog number :IF9302

The glutathione S-transferase (GST) pull-down technique has become an invaluable tool for the life scientist interested in protein chemistry.

The basic pull-down assay is an in vitro technique that consists of a GST-tagged bait protein (the

researcher’s protein of interest) that can be used to identify putative binding partner(s) (the prey). The bait protein, purified from an appropriate expression system (e.g., Escherichia coli or baculovirus-infected insect cells), is immobilized to glutathione beads. The bait serves as the secondary affinity support for confirming a previously suspected protein partner or for identifying new protein partners to the bait. Prey protein can be obtained from multiple sources, including previously purified proteins, cell lysates or in vitro transcription/translation reactions.

Protein:protein interactions can be visualized by SDS-PAGE and associated detection methods depending on the sensitivity requirements of the interacting proteins. These methods include coomassie, silver, zinc staining and Western blotting.

Overview

Tested applications: GST Pull-Down Assay.

Optimal amount should be determined by the end user.

Product Picture

Specificity: Binding GST-Tagged Fusion Protein.

Properties

Form: Magnetic Beads in Suspension (20% slurry), supplied in phosphate buffered saline (PBS) pH 7.4, with 0.02% sodium azide as preservatives.

Storage instruction: Store at 4°C. This product is stable for 12 months. Do not freeze, dry or centrifuge magnetic beads. This may cause irreversible aggregation and decreased binding capacity.

Bead Diameter:	40 µm
Concentration:	20% slurry
Binding Capacity:	5 to10 mg GST fusion protein per 1 mL settled beads (5mL 20% slurry).

Applications

Application Image

Protocols: Procedure Summary for GST Pull-Down Assay

I. Preparation of Bait Protein

Protocol 1: Bait from E. coli Expression Systems

Protocol 2: Bait from Insect Cell Expression Systems

Protocol 3: Bait from Previously Purified Protein

II. Immobilization of Bait Protein

A. Preparation of Glutathione beads

B. Immobilize Bait Protein

III. Preparation of Prey Protein

Protocol 1: Prey Protein from E. coli or Baculovirus-Infected Insect Cell Lysate

Protocol 2: Prey Protein from Mammalian Cell Lysate

Protocol 3: Prey from Previously Purified Protein

Protocol 4: Prey Protein from In vitro Transcription/Translation Reaction

IV. Capture of Prey Protein

V. Elution of Bait-Prey

VI. SDS PAGE Gel Analysis or WB Analysis

Glutathione Magnetic Beads for GST Pull-Down Assay

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