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

Cat. No.: C1456040

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

Overview

UltraBio™ Carboxyl Magnetic Beads (1 μm) are magnetic microspheres with high saturation magnetization, surface-functionalized with carboxyl groups. The carboxyl groups on these beads enable covalent coupling of biological ligands (such as peptides, proteins, antibodies, and oligonucleotides) to the microsphere surface via specific chemical reagents (e.g., EDC). This facilitates rapid isolation and enrichment of target molecules from complex samples, making them a vital tool in medical and molecular biology research. 

Parameter	Specification
Matrix	Polymer Magnetic Microspheres
Ligand	Carboxyl
Bead Concentration	10 mg/mL
Particle Size	1 μm
Coupling Capacity	>10μg IgG/mg beads
Storage	2-8°C (in Deionized water, 0.05% Kv300)

Instructions for Use:

I. Buffer Preparation

• Activation Solution: 50 mM MES, pH 5.0

• EDC Solution: 10 mg/mL in 50 mM MES, pH 5.0 (Prepare fresh)

• Blocking Solution: 50 mM Tris, pH 7.4 OR 50 mM Ethanolamine, pH 8.0

• Wash Solution: 1× PBS, pH 7.4 containing 0.1% Tween-20 or Triton X-100

• Storage Solution: Deionized water, 0.05% Kv300

II. Bead Activation

1. Transfer 200 µL of UltraBio™ Carboxyl Magnetic Beads (1 μm) to a microcentrifuge tube. Vortex-mix thoroughly.

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

   • Note: Avoid aspirating beads. Applies throughout.

3. Remove the tube from the rack. Add an equal volume of Activation Solution. 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 Step 3 & 4 once more (1 wash total with Activation Solution).

6. Add 200 µL of freshly prepared EDC Solution (10 mg/mL). Mix thoroughly.

7. Incubate at room temperature for 30 minutes. Ensure the beads remain uniformly suspended to maximize activation efficiency.

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

9. Wash the beads twice rapidly with pre-chilled Deionized Water followed by two rapid washes with Activation Solution (Steps 3 & 4). Minimize time to prevent hydrolysis of activated groups.

III. Ligand Coupling

1. Add 200 µL of ligand solution (e.g., antibody/protein at 1-5 mg/mL in Activation Solution) to the activated beads. Mix thoroughly.

2. Incubate with end-over-end rotation at room temperature overnight (12-16 hours). Ensure the beads remain uniformly suspended.

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

4. Add 500 µL Blocking Solution. Incubate at room temperature for 30 minutes.

5. Aspirate the blocking solution. Wash beads twice with Wash Solution, followed by two washes with 1× PBS, pH 7.4.

6. Resuspend the coupled beads in Storage Solution for immediate use or store at 2-8°C for long-term storage.

IV. 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

Specifications & Purity

BioReagent, 10 mg/mL

Stability And Storage

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

Storage

Store at 2-8°C, Do not freeze

Shipped In

Wet ice, Do not freeze

This product requires cold chain shipping. Ground and other economy services are not available.

Grade

BioReagent