Protein Purification Magnetic Beads/Kits - Efficient Capture of Target Molecules

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June 16, 2025

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Experimental Principle

Protein purification magnetic beads/kits achieve precise capture and purification of target molecules based on specific biomolecular interactions. The surface of the magnetic beads is modified with high-affinity ligands (such as antibodies, streptavidin, His-tag binding groups, etc.), which can specifically bind to the target protein. For example, antibody-coupled magnetic beads selectively adsorb the target protein through the antigen-antibody binding principle, while streptavidin magnetic beads utilize the high affinity of biotin-avidin to capture biotinylated molecules.

After binding, magnetic separation technology is used to quickly separate the magnetic bead-target molecule complex from impurities, followed by washing to remove non-specific adsorption. Finally, the high-purity target product is released by changing the buffer conditions (such as pH adjustment, competitive elution). This technology has the advantages of high efficiency, gentleness, and automation, and is suitable for the purification of trace targets in complex samples (such as serum, cell lysates).

Experimental Procedure (Using Antibody-Conjugated Magnetic Beads as an Example)

1、Sample Pretreatment

Centrifugal Clarification: Centrifuge the sample (such as serum) at 3000g for 10 minutes to remove cellular debris or particulate matter.

Lysis (Optional): If the target is intracellular components, treat the sample with a lysis buffer containing protease inhibitors, sonicate to disrupt cells, and then centrifuge to collect the supernatant.

2、Magnetic Bead Incubation

Magnetic Bead Activation: Vortex mix the magnetic bead suspension, take an appropriate amount of magnetic beads into a centrifuge tube, and wash twice with pre-chilled PBS to remove the storage solution.

Binding Target Molecules: Mix the pretreated sample with the magnetic beads and incubate at room temperature with rotation for 30 minutes to ensure full contact.

3、Magnetic Separation and Washing

Magnetic Absorption to Remove Supernatant: Place the centrifuge tube on a magnetic rack and let it stand for 2-3 minutes until the solution is clear, then aspirate the supernatant.

Washing to Remove Impurities: Add washing buffer (such as PBS containing 0.1% Tween-20), gently resuspend the magnetic beads, and repeat the magnetic absorption step 2-3 times to remove non-specific bindings.

4、Target Molecule Elution

Competitive or Conditional Elution:

Acidic Elution: Add a low-pH buffer (such as glycine-HCl with pH 2.8), incubate for 5 minutes, neutralize immediately, and transfer the supernatant to protect the activity of the target.

Biotin Competition: For streptavidin magnetic beads, add an excess of free biotin to competitively release the target molecules.

Magnetic Bead Regeneration (Optional): Some kits support magnetic bead regeneration, which can be reused after treatment with deionized water and regeneration buffer.

Total Time: Approximately 60 minutes, with inter-batch variation <5%, and recovery rate can reach over 90%.

Product Advantages

Release microspheres to break free from the limitations of sample volume, reagent flow rate, column pressure, and elution volume.
Simultaneously achieve solid-liquid and liquid-liquid separation, simplifying steps, reducing loss, and saving time.
Break through the technical bottlenecks in the purification of secreted and inclusion body proteins.
Reduce dependence on instruments and equipment, lowering maintenance and service costs.
Stable operation, facilitating high-throughput and large-scale protein purification.
Stable product performance with minimal batch-to-batch variation.
Achieve high yield and high purity of the target protein.

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Absin provides antibodies, proteins, ELISA kits, cell culture, detection kits, and other research reagents. If you have any product needs, please contact us.

Absin Bioscience Inc.
Email: worldwide@absin.net