Surface Activated Dynabeads

Transcription

1 Surface Activated

2 Surface Activated A variety of ways to bind your ligand Manual or automated handling Protein separation technology Superior reproducibility All-in-one-tube protocols Easy handling and no sample loss Introduction are superparamagnetic monosized polymer spheres. They are magnetic only when placed in a magnetic field, with no magnetic remanence when the magnetic field is removed. are composed of highly cross-linked polystyrene with magnetic material precipitated in evenly distributed pores. The ferrous compound is sealed inside the bead by an additional layer of polymer. Chemical groups are then introduced onto the bead surface to facilitate binding of almost any ligand, and subsequent easy and convenient isolation of the bound target (fig. 1). Unique Monodispersity The unique monodispersity and uniformity of the provide optimal and reproducible reaction kinetics, and a rapid and efficient binding of target molecules. are easily dispersed and handle almost like a liquid. In addition, chemical agglutination and nonspecific binding are negligible as compared with irregularly shaped particles. Hydrophobic or Hydrophilic Dynal Biotech provides surface activated with both hydrophobic and hydrophilic characteristics: The hydrophilic beads allow for gentle coupling of ligands to the surface. The functional ac tivit y of enz y mes and labile proteins is therefore intact after immobilisation. T he hydrophilic proper ties of the beads ensure optimal dispersion in suspension. Very low nonspecific binding of proteins, dyes etc. is observed with the ph neutral M-270 Epoxy beads, thus the requirement for blocking agents is reduced. The functional groups of the M-270 Amine and Carboxylic Acid beads allow further introduction of a large variety of alternative reactive groups by using commercially available cross-linkers. A hydrophobic surface is an excellent choice for coupling of antibodies for affinity purification of organelles or proteins. The hydrophobic Fc region of the antibody is adsorbed to the hydrophobic bead surface, followed by a rapid covalent bond formation. The orientation of the antibody is optimal, with the Fab regions facing outwards. The resultant yield of target protein or organelle fractionation is elevated. Small or Large, Automation or Manual Handling Dynal Biotech offers surface activated of various sizes. For work with proteins, nucleic acids or similar materials, we recommend the 1.0 µm (the MyOne TM series) or the 2.8 µm (the M-280 and M-270 series). The larger 4.5 µm beads are well suited to isolating larger entities such as organelles or cells. Due to their size and high iron content they exert a strong pull to the magnet, even in viscous samples, yet still retain gentle handling of the bound target. The are easy to handle manually using simple magnet racks for the isolation of target. MyOne TM are ideal for isolation of smaller entities such as proteins and bacteriophages. In addition, the 1.0 µm beads have a large surface area, high capacity, efficient magnetic pull and excellent properties for automation. The MyOne TM products are tailor-made for use in automated protocols where high throughput is crucial. Dynal Biotech has developed suitable protocols for several of the robotic workstations available on the market. Specific Considerations To obtain the best possible result, it is important to choose the correct product for your specific application. The table on page 5 will help you decide which product to use. The choice will first of all depend on the nature of the ligand. A successful coupling of a ligand to tosylactivated beads requires primary amino- or sulphydryl groups. The orientation of the active site of the ligand must also be taken into consideration. Hydrophobic beads facilitate interactions between the beads and the hydrophobic parts of the proteins, whereas the hydrophilic beads are suitable when interactions between the beads and the hydrophilic parts of the proteins are desired. If the ligand is a labile peptide or protein, the best choice is to use beads with epoxy groups allowing for coupling down to 4 C. Carboxylic acid beads, with which peptide bonds are formed within minutes, are a satisfactory alternative. Fig. 1 Principle of biomagnetic separation with. 2

3 Ligand Antibody Target Protein or antibody Low MW antigen 1 or peptide Tosylactivated Carboxylic Acid Amine Epoxy Subcellular Organelles 2 Bacteria Virus Phage 1 Antibody fragment Protein Phage 1 or antibody Phage 1 or carbohydrate Nucleic acid Peptide Phage 1 or antibody Carbohydrate Antibody Low MV antigen Antibody Nucleic acids, oligonucleotide, aptamer, PNA Enzyme Organic chemistry derivatisation, including introduction of new functional groups Nucleic acid binding proteins 1 DNA 1, RNA 1 or PCR amplicants 1 Substrate or target for enzyme degradation 1 Streptavidin may be the best product choice 2 M-450 Epoxy best product choice for listed ligand and target alternative choice for listed ligand and target can be used for listed ligand and target 5

4 Surface Activated : M-500 Subcellular M-280 Tosylactivated MyOne TM Tosylactivated Bead Characteristics: Slightly hydrophobic bead. Surface tosyl groups. Bead diameter 4.5 µm. Hydrophobic bead. Surface tosyl groups. Hydrophobic bead. Surface tosyl groups. Bead diameter 1.0 µm. Binding Properties: Direct covalent binding to primary amino- or sulphydryl groups in proteins and Direct covalent binding to primary amino- or sulphydryl groups in proteins and Direct covalent binding to primary amino- or sulphydryl groups in proteins and No further surface No further surface No further surface Binding overnight at neutral to high ph and 37 o C. Binding overnight at neutral to high ph and 37 o C. Binding overnight at neutral to high ph and 37 o C. Main Benefits: Five layers of epoxy coating ensures a super-smooth bead surface, perfect for EM-visualisation of captured organelles. The Fc portion is generally more hydrophobic than the Fab regions, facilitating biologically active orientation of your antibody. High capacity for isolating proteins by using an immobilised antibody. The Fc portion is generally more hydrophobic than the Fab regions, facilitating biologically active orientation of your antibody. High capacity for isolating proteins by using an immobilised antibody. The Fc portion is generally more hydrophobic than the Fab regions, facilitating biologically active orientation of your antibody. Applications and Reaction Chemistries: Organelle fractionation. EM pictures of organelles. Protein purification. Isolation of fragile cells (the smaller beads are pulled gently to the magnet, ensuring the isolation of intact and viable cells). Protein purification. Isolation of rare cells (rapid kinetics due to high number of beads). IVD IVD 3

5 M-270 Amine M-270 Carboxylic Acid MyOne TM Carboxylic Acid M-270 Epoxy Surface amino groups. Direct covalent binding through reductive amination of aldehydes. No further surface Rapid binding (less than 1 hour) at neutral to high ph and room temperature. Surface carboxylic acid groups. Covalent amide bond formation with primary amino- and sulphydryl groups in proteins and Activation through carbodiimide is required. Immediate peptide bond formation at ph 5-6 and room temperature. Surface carboxylic acid groups. Bead diameter 1.0 µm. Covalent amide bond formation with primary amino- and sulphydryl groups in proteins and Activation through carbodiimide is required. Immediate peptide bond formation at ph 5-6 and room temperature. Surface epoxy groups. Direct covalent binding to primary amino and sulphydryl functional groups in proteins and No further surface Binding overnight at neutral ph, high salt and a wide temperature range. Rapid immobilisation of carbohydrates, glycoproteins and glycolipids (e.g. lipopolysaccharides). Easy introduction of further alternative surface chemistries. 100% covalent binding. Low non-specific binding of nucleic acids. Rapid binding chemistry. 100% covalent binding. Low non-specific binding of nucleic acids. Rapid binding chemistry. The functionality of your enzyme is maintained after isolation. Very low non-specific binding of proteins is observed with these ph neutral beads. Isolation of cells, proteins or other target molecules with affinity for specific carbohydrate moieties. These beads can be used to introduce other reactive groups by coupling different cross-linking reagents. C-terminal coupling of Immobilisation of labile proteins and N-terminal coupling of IVD Immobilisation of labile proteins and N-terminal coupling of IVD NA IVD Purification of temperaturelabile proteins and active proteins such as enzymes. Immobilisation of protein A and G. Analysis of enzymatic reactions by coupling the enzyme directly onto the beads. 4

6 Ordering Information Product Concentration Volume Product No. M-270 Amine Hydrophilic beads with amino groups M-450 Epoxy Hydrophobic beads with epoxy goups. 5 ml M-270 Epoxy Hydrophilic beads with epoxy goups. 60 mg 300 mg freeze-dried freeze-dried M-450 Tosylactivated Hydrophobic beads with tosyl group M-280 Tosylactivated Hydrophobic beads with tosyl group. 100 mg/ml MyOne Tosylactivated Hydrophobic beads with tosyl group. 100 mg/ml M-270 Carboxylic Acid Hydrophilic beads with carboxylic acid groups MyOne Carboxylic Acid Hydrophilic beads with carboxylic acid groups. 10 mg/ml 100 ml M-500 Subcellular Slightly hydrophobic beads with tosyl groups Dynal MPC -S Unit Dynal MPC -96S Unit products for cell separation and other applications are available at. If you need assistance in choosing the right product please contact our Technical Centre. Copyright 2005 Dynal Biotech ASA, Norway. Printed All rights reserved. Dynal, and Dynal MPC are registered trademarks of Dynal Biotech ASA. The products are covered by several international patents and patent applications. Dynal Biotech is an ISO 9001 certified company. Dynal Biotech will not be responsible for violations or patent infringements that may occur with the use of our products. Dynal Biotech ASA, Norway, tel: Dynal Biotech LLC, USA, tel: Dynal Biotech GmbH, Germany, tel: Dynal Biotech Ltd, UK, tel: Dynal Biotech S.A, France, tel: Dynal Biotech Pty Ltd, Australia, tel: Dynal Biotech Beijing Ltd, tel: Nihon Dynal K.K., Japan, tel: MOL.B