UltraBio™ Epoxy Magnetic Beads (1μm) - BioReagent, 10 mg/mL

Cat. No.: E1456039

GRADE & PURITY BioReagent, 10 mg/mL

Storage

Store at 2-8°C,Do not freeze

Shipped In

Wet ice,Do not freeze

Application

Nucleic Acid Extraction and Purification, Protein Separation and Purification

Descripción general

UltraBio™ Epoxy Magnetic Beads (1 μm) are epoxy-activated, superparamagnetic magnetic particles. As a widely utilized functional biological magnetic bead, they are primarily used for bioanalytical detection. These beads enable direct coupling with proteins and samples containing amino, thiol, or hydroxyl groups. This pre-activated medium can be customized to prepare specialized affinity media for the rapid and efficient one-step purification of target substances from complex systems. 

Parameter	Specification
Matrix	Polymer Magnetic Microspheres
Ligand	Epoxy
Bead Concentration	10 mg/mL
Particle Size	1 μm
Coupling Capacity	>10μg IgG/mg beads
Storage	2-8°C (in 100% Isopropanol)

Instructions for Use:

I. Buffer Preparation

• Coupling Buffer: 0.1 M Na₂CO₃, pH 9.5

• Blocking Solution: 1 M Ethanolamine, pH 8.0

• Wash Buffer 1: 0.1 M Sodium Acetate-Acetic Acid, 0.5 M NaCl, pH 3.0

• Wash Buffer 2: 0.1 M Tris-HCl, 0.5 M NaCl, pH 8.0

• Storage Solution: PBS, 0.01% Tween-20, 0.02% NaN₃

  • *Note: Dissolve the sample in Coupling Buffer to a concentration of ~5-10 mg/mL. Coupling buffers (e.g., carbonate, phosphate) should lack primary amine groups.*

II. Sample Coupling

1. Invert the UltraBio™ Epoxy-Activated Magnetic Beads (1 μm) vial repeatedly to ensure thorough mixing (bead concentration: 10 mg/mL). Transfer the desired volume of bead suspension to a microcentrifuge tube.

2. Place the tube on a magnetic rack for ~1 minute until the solution clears. Aspirate and discard the supernatant.

3. Remove the tube from the rack. Add deionized water. Pipette mix 5-10 times to resuspend the beads.

4. Place the tube on the magnetic rack for ~1 minute. Aspirate and discard the supernatant.

5. Repeat Steps 3 & 4 once more (2 water washes total).

6. Wash the beads once with Coupling Buffer (Steps 3 & 4).

7. Add the prepared sample solution to the washed beads. The final bead concentration in the sample mixture should be ~30-50 mg/mL. Mix by inversion.

8. Incubate the tube with end-over-end rotation at room temperature for 24-48 hours. Ensure the beads remain uniformly suspended during incubation to maximize coupling efficiency.

9. Place the tube on the magnetic rack for ~1 minute. Aspirate and save the supernatant for coupling efficiency analysis if needed.

10. Wash the beads sequentially with:

    • Deionized Water

    • Wash Buffer 1

    • Deionized Water

    • Wash Buffer 2

    • Deionized Water
      Perform magnetic separation and aspirate supernatant after each wash. Repeat this alternating wash cycle once more (2 cycles total).

11. Resuspend the coupled beads in an equal volume of Storage Solution. Store at 2-8°C.

III. Purification Procedure (Example: Antigen Purification using Antibody-Coupled Beads)

1. Buffer Preparation:

   • Filter all water and buffers (0.22 μm or 0.45 μm) before use.

   • Equilibration/Wash Buffer: 0.15 M NaCl, 20 mM Na₂HPO₄, pH 7.0

   • Elution Buffer: 0.1 M Glycine, pH 3.0

   • Neutralization Buffer: 1 M Tris-HCl, pH 8.5

2. Bead Pre-equilibration:

   • Resuspend coupled beads thoroughly. Transfer the calculated volume (e.g., 100 μL) to a microcentrifuge tube.

   • Place on magnetic rack for ~1 min until solution clears. Aspirate supernatant.

   • Remove tube from rack. Add an equal volume (e.g., 100 μL) of Equilibration Buffer. Pipette mix 5 times.

   • Place on magnetic rack for ~1 min. Aspirate supernatant.

   • Repeat equilibration wash twice (3 washes total).

3. Sample Binding:

   • Add sample solution to the pre-equilibrated beads. Vortex-mix briefly.

   • Incubate with end-over-end rotation (or gentle manual tube inversion) at room temperature for ≥30 min (adjust based on binding efficiency).

   • Place on magnetic rack for ~1 min. Aspirate and discard supernatant.

4. Washing:

   • Add 5x initial bead volume (e.g., 500 μL) of Wash Buffer. Vortex-mix to resuspend.

   • Incubate with mixing for 1-2 min.

   • Place on magnetic rack for ~1 min. Aspirate and discard supernatant.

   • Repeat wash twice (3 washes total).

5. Elution:

   • Add 3-5x initial bead volume (e.g., 300-500 μL) of Elution Buffer. Pipette mix 5 times.

   • Incubate with end-over-end rotation (or gentle manual tube inversion) at room temperature for 5-10 min.

   • Place on magnetic rack for ~1 min. Transfer the clear supernatant (elution fraction) to a clean tube. This contains the target protein.

6. Neutralization:

   • Immediately add 1/10th volume of the elution fraction of Neutralization Buffer to the elution fraction. Mix thoroughly to adjust pH to 7.0-8.0.

7. Bead Regeneration & Storage:

   • Add 1 mL Elution Buffer to the beads. Vortex-mix. Place on magnetic rack for ~1 min. Aspirate supernatant.

   • Repeat elution wash once (2 washes total).

   • Add 1 mL Equilibration Buffer. Vortex-mix. Place on magnetic rack for ~1 min. Aspirate supernatant.

   • Resuspend beads in Storage Solution (Section I) at 10 mg/mL. Store at 2-8°C.

Specifications

Especificaciones y pureza

BioReagent, 10 mg/mL

Estabilidad y almacenamiento

Store at 2-8℃ long term (24 months). Do not freeze.

Condiciones de almacenamiento de almacenamiento

Store at 2-8°C, Do not freeze

Enviado en

Wet ice, Do not freeze

Este producto requiere envío en cadena de frío. Los servicios terrestres y otros servicios económicos no están disponibles.

Grado

BioReagent