MagSIGNAL-COOH 300 beads

Details for Product No. ABIN1721151, Supplier: Log in to see
Antigen
Application
Immunoassay (IA), Separation (Sep)
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Purpose Superparamagnetic particles of 300 nm with a carboxyl modified surface. Intended for immobilization of antibodies or proteins with carbodiimide coupling chemistry with NH2-containing molecules, and use as a detection label in magnetic immunoassays or as solid support phase in immunoassays with other readout techniques.
Components Magnetic beads with carboxyl activated surface
Material not included Magnet for separation of beads
Vortex and shaker
1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC)
N-hydroxysuccinimide (NHS)
MES Buffer
PBS Buffer
Blocking Buffer
PBST Buffer
Storing Buffer
ProductDetails: Bead Ligand Carboxyl modified surface
ProductDetails: Bead Matrix Superparamagnetic Silica particles
ProductDetails: Bead Size Bead size: 300 nm
Application Notes MagSIGNAL beads consist of many ferromagnetic grains with random oriented magnetic moments, encapsulated in a non-magnetic silica shell. The size of the ferromagnetic grains is 10-15 nm. Due to the small size and random orientation of the magnetic moments, the beads behave superparamagnetically and the magnetic moment without a magnetic field is 0 (demonstrated in figure 1 below).
Comment

Magnetic immunoassays (MIA) use magnetic beads as labels instead of conventional enzymes (ELISA), fluorophores or luminescent molecules. These assays involve the specific binding of an antibody with a conjugated magnetic label to its antigen. The presence of magnetic beads is then detected by a magnetic reader (magnetometer) which measures the magnetic field change induced by the beads. The signal measured by the magnetometer is proportional to the analyte (virus, toxin, bacteria, cardiac marker,etc.) quantity in the initial sample.
Magnetic beads are not affected by reagent chemistry or photobleaching and are therefore stable over time. Because measurements are based on nonlinear magnetic signals, the contribution of linear magnetic signals from materials such as sample matrix and consumables are eliminated. The technology makes magnetic immunoassays possible in a variety of formats such as conventional lateral flow test (replacing gold labels with magnetic labels), microfluidic applications and biochips.

Protocol MagSIGNAL are superparamagnetic beads with a mean diameter of 300 nm, which can be used as a detection label in magnetic immunoassays, or as solid support phase in immunoassays with other readout techniques.
MagSIGNAL-COOH 300 is intended for covalent antibody coupling for specific analyte detection. Other applications include protein coupling in combination with the use of a specific antibody - for instance, streptavidin coupling and a biotinylated antibody.
Reagent Preparation

MES Buffer: 0.1 M 2-(N-Morpholino)ethanesulfonic acid (MES), pH 5.5
PBS Buffer: 0.01 M PBS pH 7.4
Blocking Buffer: 0.1 M Tris Hydroxymethylaminoethane (TRIS buffer)
PBST Buffer: 0.01 M PBS pH 7.4 with 0.05% Tween 20
Storing B

Assay Procedure

For covalent coupling of amino- group containing ligands such as antibodies, proteins or low molecular substances to MagSIGNAL-COOH, the carbodiimide method can be applied.
Carbodiimides react with the terminal carboxylate-groups from the magnetic beads to highly reactive O-acylisourea derivatives and react readily with amino-groups of the ligands, as presented in Figure 1 below.

Coupling protocol
The following protocol describes the coupling of biomolecules on 1 mL (10 mg) beads. It can be scaled up by adjusting volumes of required reagents.
1.Resuspend MagSIGNAL-COOH 300 by vortexing. Transfer 1 mL from the storage bottle to a reaction container.
2.Wash the beads:Place the reaction container in a magnetic separator and wait until the separation is completed. Discard the buffer and resuspend in 1 mL MES Buffer. Repeat this step once more.
3.Resuspend the beads in 0.50 ml MES Buffer containing 20 mg EDC and 20 mg NHS (freshly prepared!) and incubate on a shaker for 15 minutes at room temperature.
4.Place the reaction container in a magnetic separator, wait until the separation is completed, and discard the supernatant. Resuspend the beads in 1 mL MES Buffer.
5.Repeat step 4 once more. The beads now contain activated COOH groups that can bind proteins and other amine containing molecules.
6.Place the reaction container in a magnetic separator, wait until the separation is completed, and discard the supernatant.
7.Add ligands to the activated beads and add PBS Buffer to a final coupling volume of 0.5 mL. Incubate on a shaker for 2 hours at room temperature.
8.Place the reaction container in a magnetic separator, wait until the separation is completed, and discard the supernatant.
9.Wash the beads 3 x with 1 mL PBST Buffer, and discard the supernatant
10.Resuspend the particles in Blocking Buffer. Incubate on a shaker for 2 hours at room temperature.
11.Wash the particles 3 x with 1 mL PBST Buffer and 3 x with 1 mL PBS Buffer. Discard the supernatant.
12.Resuspend the beads in Storing Buffer and store at 2-8 °C.

Restrictions For Research Use only
Format Liquid
Concentration 10 mg/mL
Buffer PBS (pH 7.4), 0.05% Sodium azide
Preservative Sodium azide
Precaution of Use This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Handling Advice Store in well closed vial and in upright position to prevent drying, this may result in a decrease of activity.
Do not freeze the product!
Vortex well before use. Wash the beads to remove preservatives that could interfere with your appl
Storage 4 °C
Expiry Date 12 months
Supplier Images
 image for MagSIGNAL-COOH 300 beads (ABIN1721151) Carbodiimide coupling method for MagSIGNAL-300 COOH
 image for MagSIGNAL-COOH 300 beads (ABIN1721151) MagSIGNAL-COOH 300 beads (Image 2)
 image for MagSIGNAL-COOH 300 beads (ABIN1721151) Magnetization curve of MagSIGNAL (sample amount: 15,4 mg, normalized moment at 700 Oe...