Carboxyl beads

MACS (ImpetiCbead, IMB referred beads), in recent years developed a new immunological techniques, the unique height advantage and immunological reaction curing agents will specifically bind to one to immunological basis, penetrated into all areas of pathology, physiology, pharmacology, microbiology, biochemistry and molecular genetics, which has been more widely used in immunoassays, cell separation, purification of biological macromolecules and molecular biology.

Principle:

Using core - shell synthetic polymer synthesis surface four superparamagnetic iron oxide-containing polymer cover, good stability, can be post-labeled substance, the use of these substances surface features such as an amino group, a carboxyl group, a thiol group, etc. antibody covalently or non-covalent coupling can be used to bind the corresponding antigen, so that under the applied magnetic field to attract do directional movement, so as to achieve the separation, detection, purification of target genes, proteins, cells, microorganisms and the like.

Main applications:

1, Gene purification: use of the silicon bead surface may be reversible under certain conditions of the adsorption and dissociation characteristics of DNA / RNA can be used to purify the gene, genes isolated object, such as genomic DNA, material DNA, viral DNA, whole blood genomic DNA, RNA and the like.

2, protein purification: the use of a carboxyl group, an amino group, streptomycin, avidin, a protein sulfhydryl cross-linking corresponding antibodies can specifically and effectively isolated to the corresponding protein.

3, cell purification: this area has become a major application of MACS.

4, microbial enrichment: the use of microbial corresponding antibody beads can efficiently be reversible enrichment corresponding microorganisms.

Features:

YR-NH2-01

1. The amino abundant:> 150μmol / g;

2. Operating performance: beads uniformly dispersed superparamagnetic, magnetic response time <30s;

3. Inter-batch stability and good repeatability: uniform particle size, polydispersity index <0.2, was monodisperse;

YR-NH2-02

1. Amino-rich content: ~ 1000μmol / g;

2. Operating performance: beads uniformly dispersed superparamagnetic, magnetic response time <10s;

3. Inter-batch stability and good reproducibility, carboxyl content between batch CV <5%;

The goal of high volume substances binding, nonspecific adsorption is low: dedicated to separation and purification field.

Product advantages:

1. rich binding sites, strengthen specific binding ligand.

2. superparamagnetic and high magnetic response, time-saving operation.

3. Good dispersion and resuspended, improve ease of operation.

4. good physical and chemical stability, repeatability guarantee results.

Precautions:

1. freezing, drying and centrifugation action will cause beads reunion, is not easy to resuspended and dispersed, and affect the chemical activity of bead surface functional groups.

2. Before using this product, be sure to thoroughly shaken or ultrasound make beads to maintain a uniform suspension.

3. The use of the process according to the needs, with purified water or a buffer solution magnetic beads were washed 2 to 3 times to remove the preservation solution in ethanol.

4. This product needs and supporting the use of magnetic separation equipment.

5. In order to ensure the best results, please use the appropriate ligand covalent coupling reaction.

6. remain stable at 2 ~ 8 C, the shelf life of 2 years.

7. This product is for research use

The method of magnetic beads conjugated with biological molecules (refer to protein A as an example):

A. Bead surface carboxyl activating

1. After mixing beads, magnetic beads to take 100μL 1mL microcentrifuge tube, magnetic removal of the supernatant solution with 200μL PBS (50mM PBS, pH 7.4) were magnetically separated and washed 2 times, and then the supernatant was removed;

2. rapidly added a solution of freshly prepared 100μL EDC (10mg / mL, the above solution as a dispersant MEST) and 100μL NHS (10mg / mL, the above solution as a dispersant MEST) was added to the centrifuge tube containing the beads, Swirl bead mix so full suspension, 25 C activation 30min, maintaining the period of suspension beads (which can be reversed with a vertical mixer and mix); After the above steps, carboxyl bead surface has been activated, with belt There are primary amino biological ligand is covalently coupled. (Activation state for too long preservation, the proposed coupling immediately)

B. Covalently coupled to magnetic beads and the biological ligand

1. The magnetic separation to remove the supernatant, was added 50μg ~ 200μg biological ligands (appropriate amount and concentration need to be optimized according to specific experiments, maintaining the solution pH≈8.0, 0.05% Tween 20 may be added to improve the dispersibility of the beads), gently mix;

2. 25 C coupling 2h, or 25 C coupling 1h after placing 4 C stand overnight, keeping the beads in suspension (which can be reversed with a vertical mixer mix) during the coupling;

3. The centrifuge tube was placed in the magnetic separation rack magnetic separation to remove supernatant, added 200μL PBST solution (pH 7.2, containing 1% BSA) resuspend beads (ultrasound may be required), 25 C reactor 1h closed bead surface unreacted activated carboxyl group, for the period to keep the beads in suspension (which can be reversed with a vertical mixer and mix);

The centrifuge tube was placed on a magnetic separator after the magnetic separation to remove the supernatant, each was treated with 200μL PBS (pH 7.2) or saving solution was washed 3 times, resuspended in preservation solution (which can be determined according to the need to preserve the solution amount, in order to adjust the concentration of conjugated ligand beads), stored at 4 C. If the immobilized biological ligand stability, may be added 0.02% (W / V) in the preservation solution of sodium azide (NaN) as a bacteriostatic agent.