Streptavidin MagPoly Beads

Streptavidin MagPoly Beads 

1. Product Description

Streptavidin MagPoly Beads is an affinity chromatography Magnetic beads designed for easy, one-step purification of biotin or biotinylated proteins, antibodies, and other substances through the interaction between biotin and ligand. The streptavidin ligand is coupled to Magnetic beads.

Table 1. Characteristics of Streptavidin MagPoly Beads

2. Purification Procedure

The protocol uses 1mg Streptavidin MagPoly Beads, but this may be scaled up or down as required.

• Preparation of the Magnetic Beads
• Completely resuspend the beads by shaking or vortexing the vial.
• Transfer 100 μl Streptavidin MagPoly Beadsto a new tube.
• Place the tube on a magnetic separation rack to collect the beads at tube wall. Remove and discard the supernatant.
• Add 0.5 ml selected washing buffer to the tube and invert the tube several times to mix. Use the magnetic separation rack to collect the beads and discard the supernatant. Repeat this step twice.

Recommended washing buffers:

– nucleic acid applications: TES Buffer

– antibody/protein applications: PBS Buffer, pH 7.4

• Method for Immobilization of Biotinylated Molecules
  • Additional Materials Required

Biotinylated sample in solution:

Binding/Wash Buffer: Nucleic acid applications: TES Buffer;

Protein/antibody applications: PBS, pH 7.4.

• Procedure
• Resuspend the beads in 100 μl Binding/Wash Buffer.
• Add biotinylated sample to the beads prepared from step 2.2.1 and gently invert the tube to mix.
• Incubate the tube at room temperature with mixing (on a shaker or rotator) for one hour.
• Use the magnetic separation rack to collect the beads and discard the supernatant. If desired, keep the supernatant for analysis.
• Add 1 ml Binding/Wash Buffer to the tube and mix well, use the magnetic separation rack to collect the beads and discard the supernatant. Repeat the wash step three times.
• Resuspend to desired concentration in a suitable buffer for your downstream use.
• Method for Purifying Antigens
  • Additional Materials Required

Biotinylated antibody: Use approximately 2-3 mg of biotinylated antibody/ml settled Streptavidin MagPoly Beads

Binding/Wash Buffer: 0.1 M phosphate, 0.15 M NaCl, pH 7.0

Elution Buffer: 0.1 M glycine HCl, pH 2.5 - 2.8

Neutralization buffer: 1 M Tris HCl, pH 8.5

• Procedure

• Resuspend the beads in 100 μl Binding/Wash Buffer.
• Add biotinylated antibody solution to the beads prepared from step 2.3.1 and gently invert tube to mix.
• Incubate the tube at room temperature with mixing (on a shaker or rotator) for one hour.
• Use the magnetic separation rack to collect the beads and discard the supernatant. If desired, keep the supernatant for analysis.
• Add 1 ml Binding/Wash Buffer to the tube and mix well, use the magnetic separation rack to collect the beads and discard the supernatant. Repeat the wash step three times.
• Resuspend the antibody bound beads in 100 μl Binding/Wash Buffer.
• Add antigen sample to the tube and gently invert tube to mix. Incubate at room temperature for 30 minutes to overnight at 4°C.
• Wash the beads with 1ml Binding/Wash Buffer. Use the magnetic separation rack to collect the beads and discard the supernatant. Repeat the wash step three times.
• Add 100 μl Elution Buffer to the tube. Mix well and incubate for five minutes at room temperature with occasional mixing.
• Use the magnetic separation rack to collect the beads and save the supernatant containing target antigen.
• To neutralize the low pH, add 10μl neutralization buffer to each 100 μl eluate.

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