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Azide group functional magnetic microparticles (PuriMag G-N3 Beads)

2025/6/17 viewers
  • BrandPuriMag
  • TypePuriMag G Series
  • order
Introduction

Ordering information

Name

Cat. No.

Vol.

Scheme

G-N3

PMG012-2

2 ml


1. Overview

Click Chemistry describes rapid, selective "click" reactions between paired functional groups in mild aqueous solutions. This concept has evolved into a convenient, versatile, and reliable two-step coupling procedure for molecules A and B, widely adopted in biosciences, drug discovery, and materials science.

PuriMag? G-N?, Azide-Functionalized Magnetic Nanoparticles are uniform, polymer-coated superparamagnetic nanoparticles. Their hydrophilic surface ensures excellent dispersibilitylow non-specific adsorption, and easy handling in diverse buffers. The beads feature a high-density surface coating of azide active groups, enabling covalent capture of functionalized proteins via:

  • Cu(I)-catalyzed Azide-Alkyne Cycloaddition (CuAAC)

  • Cu(I)-free Azide-DBCO Click Chemistry

Workflow Requirements:

  • Target proteins must be metabolically, enzymatically, or chemically tagged with:

    • Alkyne groups (for CuAAC)

    • DBCO groups (for Cu(I)-free reactions)

  • Post-coupling, rigorous washing eliminates non-specifically bound proteins.

  • Unique surface chemistry ensures low background binding without surfactants.

Features & Advantages:
High binding capacity
Rapid, efficient coupling
Hydrophilic long-arm spacers minimize steric hindrance/non-specific binding
Charge-neutral surface post-coupling
Stable covalent bonds with minimal ligand leakage
Low non-specific binding
? Capacity: 1–20 mg protein or 0.1–2 mg peptide per gram beads

Caution: Azide beads are unstable in organic solvents and high urea concentrations.

Schematic Diagram of Bead Coupling Mechanism:


2. product description

Product Specifications

Description

Polymer coated Fe3O4 nanoparticles

Particle Size

200 nm

Number of Beads

~1.7×1010 beads/mg

Matrix

Proprietary polymer

Functional group

Azide group

Group density

~300 μmole / g of Beads

Magnetization

60~70 EMU/g

Formulation

10mg/mL in DI water

Storage

1 year at 2~8 ℃. Do not freeze.

3. Instructions for Use

A. Required Materials
    tert-Butyl alcohol (t-BuOH): 12 mL
    Dimethylsulfoxide (DMSO): 3 mL
    Tris[(1-benzyl-1H-1,2,3-triazol-4-yl)methyl]amine (TBTA), M.W. 530.63: 2.7 mg
    Copper(II) sulfate (CuSO?), M.W. 159.61: 16 mg
    (+)-Sodium L-ascorbate, M.W. 198.11: 20 mg
    Methanol (MeOH): 4 mL

B. Coupling Method
    For screening, optimize ligand immobilization density on PuriMag? beads by testing four concentrations: 0 μM, 5 μM, 25 μM, and 125 μM.

B-1. Solution Preparation

    1.Prepare 15 mL t-BuOH/DMSO (4:1) by mixing 12 mL t-BuOH + 3 mL DMSO.

    2.Dissolve ligand in t-BuOH/DMSO to prepare 200 μL of 500 μM ligand solution.

    3.Prepare 1 mL 5 mM TBTA (2.7 mg in t-BuOH/DMSO). Dilute 10 μL to 200 μL with t-BuOH/DMSO for 250 μM TBTA.

    4.Prepare 1 mL 100 mM CuSO? (16 mg in H?O). Dilute 10 μL to 200 μL with H?O for 5 mM CuSO?.

    5.Prepare 1 mL 100 mM sodium L-ascorbate (20 mg in H?O). Dilute 10 μL to 200 μL with H?O for 5 mM solution.

    6.Prepare 8 mL t-BuOH/DMSO/H?O (1:1) by mixing 4 mL t-BuOH/DMSO + 4 mL H?O.

B-2. Ligand Immobilization

    1.Aliquot 2.5 mg azide beads into four tubes. Separate magnetically (RT), discard supernatant.

    2.Add 500 μL t-BuOH/DMSO, disperse beads. Separate (RT), discard supernatant.

    3.Repeat Step 2 twice (total 3 washes for solvent exchange).

    4.Add reaction solutions in sequence (see table below). After adding 250 μM TBTA, disperse ultrasonically. Then add remaining solutions.

Concentration (um)

0

5

25

125

Azide beads (mg)

2.5

2.5

2.5

2.5

t-BuOH/DMSO (ul)

250

240

200

0

500uM ligands (ul)

0

5

25

125

250uM TBTA (ul)

0

5

25

125

Ultrapure water (ul)

250

240

200

0

5mM CuSO4 (ul)

0

5

25

125

5mM ()-Sodium L-ascorbate (ul)

0

5

25

125

Total  (ul)

500

500

500

500

    5.React with tube mixer 16–20 h at RT.

    6.Separate magnetically (RT), discard supernatant.

    7.Add 500 μL t-BuOH/DMSO/H?O, disperse ultrasonically.

    8.Separate (RT), discard supernatant.

    9.Repeat Steps 7–8 twice (total 3 washes).

    10.Add 500 μL 50% MeOH, disperse ultrasonically.

    11.Separate (RT), discard supernatant.

    12.Repeat Steps 10–11 twice (total 3 washes).

    13.Resuspend in 100 μL 50% MeOH. Store at 4°C.

        (Ligand-immobilized bead concentration: 0.5 mg/20 μL)

        (For research use only!)




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