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Cat. No.: HY-K0227

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NHS Magnetic Beads (200nm, 10 mg/mL)

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Storage :
4℃, 2 years

Do not freeze

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Use of MCE Products

Fluorescent Dye

Strong Fluorescence Signal / Multi-Disciplinary Applications / High Fluorescence Specificity

Description & Advantages

MCE NHS Magnetic Beads (200 nm, 10 mg/mL) contain N-hydroxysuccinimide (NHS) functional groups, which react with primary amines on proteins or other molecules to form stable amide linkages，can covalently immobilize proteins for the affinity purification of antibodies, antigens and other biomolecules. Compared with the traditional carboxyl and amino magnetic beads, NHS magnetic beads do not need to be activated by EDC/NHS or glutaraldehyde. Simply dissolve the primary amine-containing biological ligands in Coupling Buffer and incubate at room temperature to covalently couple the bioconjugate to the beads, which is easy to operate, mild coupling conditions, fast and efficient. The beads coupling process must be carried out in a buffered solvent without any amine, and automated instruments are especially useful for large screening of multiple samples.

Protocol

Recommended Buffers (Not Provided)

Buffer	Composition
Wash Buffer	1 mM HCl, pre-chilled at 4°C
Binding Buffer I	0.1 M MES, pH 5.0, for biomolecules with pI < 7
Binding Buffer II	0.1 M PBS, pH 8.0, for biomolecules with pI > 7
Blocking Buffer	0.5 M ethanolamine, pH 8.3; or 0.1 M Tris-HCl, 150 mM NaCl, pH 8.5
Storage Buffer	PBS containing 0.02% (w/v) NaN₃

Operating Instructions

Coupling of Magnetic Beads to Biomolecules (For Reference Only)

The following procedure is described using 500 μL of magnetic beads, 500 μL of biomolecule sample, and a 1.5 mL EP tube as an example. The quantities may be adjusted proportionally according to experimental requirements.

1. Protein Solution Preparation

Dissolve an appropriate amount of the protein to be coupled in binding buffer to prepare a protein solution at a concentration of 0.5-2.0 mg/mL. Store the prepared solution at 4°C until use.

Note: The protein solution must not contain substances with amino groups. Previously stored proteins should first be dialyzed or desalted to completely remove substances containing primary amino groups from the original buffer. The protein should then be prepared at a concentration of 0.5-2.0 mg/mL using the binding buffer.

2. Magnetic Bead Pretreatment

Thoroughly mix the magnetic beads and transfer 500 μL into a 1.5 mL EP tube. Perform magnetic separation and discard the supernatant. Add 1 mL of pre-chilled wash buffer and vortex for 15 s. Perform magnetic separation again and discard the supernatant. Repeat the washing procedure twice.

3. Magnetic Bead Coupling

Add 500 μL of protein solution to the magnetic beads and vortex for 30 s. Place the tube on a mixer and incubate for 1-2 h at room temperature or overnight at 4°C.

Note: Keep the magnetic beads evenly suspended during incubation. During the first 30 min of the reaction, remove the tube and vortex for 15 s every 5 min. Thereafter, remove the tube and vortex for 15 s every 15 min.

4. Blocking

Perform magnetic separation and collect the supernatant. Add 1 mL of blocking buffer to resuspend the magnetic beads, vortex for 30 s, and incubate on a mixer at room temperature for 2 h.

5. Storage

Perform magnetic separation and discard the supernatant. Add 1 mL of ultrapure water to resuspend the magnetic beads and wash the coupled product three times. Add 500 μL of storage buffer to resuspend the magnetic beads and store at 4°C.