In protein engineering and molecular biology research, affinity tag systems provide essential technical support for the efficient expression and rapid purification of target proteins. Glutathione S-transferase (GST), as a fusion tag, is widely used for the expression, isolation, and functional analysis of recombinant proteins. The GST tag is characterized by strong hydrophilicity, high expression efficiency, and good solubility, which facilitates its specific binding to glutathione and enables the mild recovery of fusion proteins under physiological conditions.
To meet researchers' needs for rapid purification and automated processing of GST fusion proteins, MtoZ Biolabs has developed glutathione magnetic agarose beads that can be broadly applicable to protein lysates derived from bacteria, yeast, or mammalian sources. The beads integrates magnetic separation and affinity purification technologies, offering laboratories a convenient, efficient, and stable solution for the purification of GST fusion proteins.
Product Overview
MtoZ Biolabs' glutathione magnetic agarose beads are composed of 30 μm superparamagnetic agarose microspheres with reduced glutathione (GSH) covalently coupled to the surface, designed for the specific binding of GST fusion proteins. The magnetic beads are uniformly dispersed and resistant to aggregation, making them suitable for purifying soluble GST fusion proteins from low-expression samples. They are compatible with both manual magnetic rack operations and automated high-throughput systems.
|
Characteristics |
Description |
|
Product content |
10 mg/ml magnetic beads in specific protective buffer |
|
Beads size |
~200 nm |
|
Magnetization |
Superparamagnetic |
|
Coupled antibody |
Anti-GST mouse monoclonal antibody |
|
Isotype |
IgG2b |
|
M.W. of antibody |
Approximately 150 kDa |
|
Antibody concentration |
≥0.6 mg GST antibody per ml beads |
|
Binding capacity |
≥ 0.6 mg GST‐tagged fusion protein per ml beads |
Protocol
1. Sample Preparation
Choose an appropriate lysis method based on the type of experimental sample (e.g., cell lysate, tissue homogenate, or in vitro expression product). It is recommended to use a buffer that does not contain strong detergents to preserve protein functionality.
2. Anti-GST Magnetic Beads Pretreatment
a. Gently resuspend the Anti-GST magnetic beads using a pipette. For every 500 μL of sample, take 20 μL of bead suspension into a clean centrifuge tube, and add 1× TBS to a final volume of approximately 0.5 mL.
b. Gently resuspend the Anti-GST beads again using a pipette. Place the tube on a magnetic rack for 10 seconds and discard the supernatant. Repeat this step twice.
c. Resuspend the Anti-GST magnetic beads in 1× TBS to the original volume.
3. Protein Binding Reaction
a. Add magnetic beads and incubate. For every 500 μL of protein sample, add 20 μL of bead suspension. Place the mixture on a nutating mixer or rotary shaker and incubate at room temperature for 2 hours or overnight at 4ºC.
b. Magnetic separation. After incubation, place the tube on a magnetic rack for 10 seconds and discard the supernatant.
c. Washing. Add 500 μL of 1× TBS, gently resuspend the Anti-GST beads with a pipette. Place on a magnetic rack for 10 seconds and discard the supernatant. Repeat the washing step three times.
4. Washing Procedure
Acidic Elution Method: This is a non-denaturing method, relatively fast and efficient. The eluted protein retains its original biological activity, facilitating subsequent analytical detection.
a. Solution preparation: Acidic elution buffer (0.1 M Glycine-HCl, pH 3.0); neutralization buffer (0.5 M Tris-HCl, pH 7.4, 1.5 M NaCl).
b. For every 20 μL of original bead volume, add 100 μl of acidic elution buffer. Mix thoroughly and place on a nutating mixer or rotary shaker, incubating at room temperature for 5 minutes. Note: incubation time should not exceed 15 minutes.
c. After incubation, place the tube on a magnetic rack for 10 seconds. Transfer the supernatant to a new centrifuge tube and immediately add 10 μL of neutralization buffer, mixing appropriately.
d. To achieve maximum elution efficiency, repeat steps b and c, and combine the same sample fractions.
e. The eluted and neutralized Flag-tagged protein can be stored at 4°C for immediate use, or at -20°C or -80°C for long-term storage.
Features and Benefits
1. Uniform Magnetic Bead Structure with Excellent Dispersibility
Moderate particle size, non-aggregating, suitable for high-throughput operations and liquid handling systems.
2. High-Affinity and Elution-Resistant Ligand
Covalently immobilized glutathione offers high stability, low elution background, and excellent purity.
3. Compatible with Low-Expression Protein Recovery
Effectively captures GST fusion proteins even from diluted lysates.
4. Compatible with both Automated and Manual Workflows
Adaptable to manual magnetic racks and high-throughput automation instruments, enhancing experimental efficiency.
5. Mild Elution Conditions
Preserves protein conformational integrity, suitable for downstream functional and interaction analyses.
Applications
1. Rapid Purification of GST Fusion Protein Expression Products
Suitable for laboratory-scale protein preparation and subsequent analysis.
2. Protein Interaction Analysis and Pull-Down Assays
Used to explore biomolecular interactions such as protein-protein and protein-ligand interactions.
3. High-Throughput Screening Experiments
Enables large-scale protein enrichment or library screening when combined with automated systems.
4. Enzymatic or Pharmacological Activity Assays
Purified proteins can be used for functional validation, small molecule screening, and binding assays.
5. Enrichment and Pre-Concentration of Low-Abundance Proteins
Enhances detection sensitivity in downstream Western blotting or mass spectrometry analysis.
MtoZ Biolabs is dedicated to providing stable and efficient protein sample preparation consumables. Glutathione magnetic agarose beads combine high-affinity glutathione ligands with high-performance magnetic agarose to offer researchers a reliable and user-friendly tool for purifying GST fusion proteins. Whether for basic laboratory procedures or automated high-throughput platform development, it demonstrates excellent stability and experimental compatibility.
For product documentation, technical support, or trial requests, please contact MtoZ Biolabs.
FAQs
Q1: Are These Magnetic Beads Suitable for All Types of GST Fusion Proteins?
A1: This product is suitable for most recombinant proteins fused with GST tags at either the N-terminus or C-terminus. It is compatible with expression systems including bacteria, yeast, insect cells, and mammalian cells.
Q2: Is the Product Compatible with Automated High-Throughput Platform Operations?
A2: Yes. Glutathione magnetic agarose beads are composed of 30 μm uniform superparamagnetic agarose particles, offering excellent dispersion and magnetic responsiveness. They are particularly compatible with automated liquid handling systems (such as 96-well plate magnetic separation platforms and high-throughput magnetic bead extractors).
Q3: Can the Magnetic Beads be Reused? How Can They Be Regenerated?
A3: The beads can be reused multiple times under gentle operating conditions, provided the protein-binding sites are not contaminated or saturated. It is recommended to thoroughly remove bound proteins using a mild elution method (e.g., reduced glutathione solution), followed by proper washing (such as alternating washes with low pH and high-salt buffers). Store the beads in a preservation solution containing preservatives. The binding capacity should be verified before each reuse to ensure the reliability and reproducibility of experimental data.
