Surface Activated
Dynabeads®
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Surface Activated Dynabeads®
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•
•
A variety of ways to bind your ligand
Manual or automated handling
Protein separation technology
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Introduction
Superior reproducibility
All-in-one-tube protocols
Easy handling and no sample loss
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.
Dynabeads ® 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. Dynabeads® 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).
Small or Large, Automation or Manual Handling
Dynal Biotech offers surface activated Dynabeads® of various
sizes. For work with proteins, nucleic acids or similar materials,
we recommend the 1.0 µm (the MyOneTM 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 Dynabeads® are easy to
handle manually using simple magnet racks for the isolation
of target.
Unique Monodispersity
The unique monodispersity and uniformity
of the Dynabeads ® provide optimal and
reproducible reaction kinetics, and a rapid
and efficient binding of target molecules.
Dynabeads® are easily dispersed and
handle almost like a liquid. In addition,
chemical agglutination and nonspecific binding are negligible as
compared with irregularly shaped
particles.
Dynabeads® MyOneTM 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
MyOneTM 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.
Hydrophobic or
Hydrophilic
Dynal Biotech provides surface
activated Dynabeads ® with
both hydrophobic and hydrophilic characteristics:
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.
The hydrophilic beads allow
for gentle coupling of ligands
to the surface. The functional
ac t i v i t y o f e n z y me s an d
labile proteins is therefore
intact af ter 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 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 Dynabeads®.
2
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Ligand
Antibody
Dynabeads®
Tosylactivated
Dynabeads®
Carboxylic
Acid
Dynabeads®
Amine
Dynabeads®
Epoxy
Protein or
antibody
√√√
√
√
√√
Low MW
antigen1
or peptide
√√√
√√
√√
√√
Organelles
√√
Target
√ √2
Bacteria
√√√
Virus
√√√
√√
√√
√√
Phage1
√√√
√√
√√
√√
Antibody
fragment
Phage1 or
antibody
√√
√√√
√√√
√√
Protein
Phage1 or
carbohydrate
√√
√√
√√
√√√
Nucleic acid
√
√√√
√
√
√√
√√√
√√√
√√
√
√√√
√
√√
√√
√√
Peptide
Phage1 or
antibody
Carbohydrate
Antibody
Low MV antigen
Antibody
Nucleic acids,
Nucleic acid
oligonucleotide, binding
aptamer, PNA
proteins1
√√
Organic
chemistry
derivatisation,
including
introduction of
new functional
groups
Substrate
or target
for enzyme
degradation
√√√
√
√√√
DNA1, RNA1 or
PCR amplicants1
Enzyme
Dynabeads®
Subcellular
√√
√
√
√√√
√
√√
√√√
√√
Dynabeads® Streptavidin may be the best product choice
Dynabeads® 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
1
2
5
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Surface Activated Dynabeads®: Dynabeads® M-500
Subcellular
Dynabeads® M-280
Tosylactivated
Dynabeads® MyOneTM
Tosylactivated
Bead Characteristics:
• Slightly hydrophobic bead.
• Surface tosyl groups.
• Bead diameter 4.5 µm.
• Hydrophobic bead.
• Surface tosyl groups.
• Bead diameter 2.8 µm.
• Hydrophobic bead.
• Surface tosyl groups.
• Bead diameter 1.0 µm.
Binding Properties:
• Direct covalent binding to
primary amino- or sulphydryl
groups in proteins and
peptides.
• Direct covalent binding to
primary amino- or sulphydryl
groups in proteins and
peptides.
• Direct covalent binding to
primary amino- or sulphydryl
groups in proteins and
peptides.
• No further surface
activation required.
• No further surface
activation required.
• No further surface
activation required.
• Binding overnight at neutral
to high pH and 37oC.
• Binding overnight at neutral
to high pH and 37oC.
• Binding overnight at neutral
to high pH and 37oC.
Main Benefits:
Applications and
Reaction Chemistries:
• Five layers of epoxy coating
• High capacity for isolating
ensures a super-smooth bead
proteins by using an
surface, perfect for
immobilised antibody.
EM-visualisation of captured
organelles.
• The Fc portion is generally
more hydrophobic than
• The Fc portion is generally
the Fab regions, facilitating
more hydrophobic than
biologically active orientation
the Fab regions, facilitating
of your antibody.
biologically active orientation
of your antibody.
• High capacity for isolating
proteins by using an
immobilised antibody.
• Organelle fractionation.
• Protein purification.
• Protein purification.
• EM pictures of organelles.
• Isolation of fragile cells
(the smaller beads are
pulled gently to the magnet,
ensuring the isolation of
intact and viable cells).
• Isolation of rare cells (rapid
kinetics due to high number
of beads).
• The Fc portion is generally
more hydrophobic than
the Fab regions, facilitating
biologically active orientation
of your antibody.
• IVD
• IVD
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Dynabeads® M-270
Amine
Dynabeads® M-270
Carboxylic Acid
Dynabeads® MyOneTM
Carboxylic Acid
Dynabeads® M-270
Epoxy
• Hydrophilic bead.
• Surface amino groups.
• Bead diameter 2.8 µm.
• Hydrophilic bead.
• Surface carboxylic acid
groups.
• Bead diameter 2.8 µm.
• Hydrophilic bead.
• Surface carboxylic acid
groups.
• Bead diameter 1.0 µm.
• Hydrophilic bead.
• Surface epoxy groups.
• Bead diameter 2.8 µm.
• Direct covalent binding
through reductive amination
of aldehydes.
• Covalent amide bond
• Covalent amide bond
• Direct covalent binding to
formation with primary
formation with primary
primary amino and
amino- and sulphydryl groups
amino- and sulphydryl groups
sulphydryl functional groups
in proteins and peptides.
in proteins and peptides.
in proteins and peptides.
• No further surface
activation required.
• Activation through
carbodiimide is required.
• Activation through
carbodiimide is required.
• No further surface
activation required.
• Immediate peptide bond
formation at pH 5-6 and
room temperature.
• Immediate peptide bond
formation at pH 5-6 and
room temperature.
• Binding overnight at neutral
pH, high salt and a wide
temperature range.
• Rapid immobilisation of
carbohydrates, glycoproteins
and glycolipids
(e.g. lipopolysaccharides).
• 100% covalent binding.
• 100% covalent binding.
•
•
• The functionality of your
enzyme is maintained
after isolation.
• Easy introduction of
further alternative surface
chemistries.
• Rapid binding chemistry.
• Rapid binding chemistry.
• Isolation of cells, proteins
or other target molecules
with affinity for specific
carbohydrate moieties.
• Immobilisation of labile
proteins and peptides.
• Immobilisation of labile
proteins and peptides.
• N-terminal coupling of
peptides.
• N-terminal coupling of
peptides.
• IVD
• IVD
• Rapid binding (less than 1
hour) at neutral to high pH
and room temperature.
• These beads can be used
to introduce other reactive
groups by coupling different
cross-linking reagents.
Low non-specific binding of
nucleic acids.
Low non-specific binding of
nucleic acids.
• NA IVD
• Very low non-specific
binding of proteins is
observed with these
pH neutral beads.
• 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.
• C-terminal coupling of
peptides.
4
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Ordering Information
Product
Concentration
Volume
Product No.
Dynabeads® M-270 Amine
Hydrophilic beads with amino groups.
30 mg/ml
2 ml
10 ml
143.07
143.08
Dynabeads® M-450 Epoxy
Hydrophobic beads with epoxy goups.
30 mg/ml
5 ml
140.11
Dynabeads® M-270 Epoxy
Hydrophilic beads with epoxy goups.
60 mg
300 mg
freeze-dried
freeze-dried
143.01
143.02
Dynabeads® M-450 Tosylactivated
Hydrophobic beads with tosyl group.
30 mg/ml
10 ml
302.12
Dynabeads® M-280 Tosylactivated
Hydrophobic beads with tosyl group.
30 mg/ml
30 mg/ml
100 mg/ml
2 ml
10 ml
10 ml
142.03
142.04
301.01
Dynabeads® MyOne™ Tosylactivated
Hydrophobic beads with tosyl group.
100 mg/ml
2 ml
10 ml
655.01
655.02
Dynabeads® M-270 Carboxylic Acid
Hydrophilic beads with carboxylic acid groups.
30 mg/ml
2 ml
10 ml
143.05
143.06
Dynabeads® MyOne™ Carboxylic Acid
Hydrophilic beads with carboxylic acid groups.
10 mg/ml
2 ml
10 ml
100 ml
650.11
650.12
650.13
Dynabeads® M-500 Subcellular
Slightly hydrophobic beads with tosyl groups.
30 mg/ml
2 ml
150.01
Dynal MPC®-S
Unit
120.20
Dynal MPC®-96S
Unit
120.27
Dynabeads® products for cell separation and other applications are available at www.dynalbiotech.com.
If you need assistance in choosing the right product please contact our Technical Centre.
©
Copyright 2005 Dynal Biotech ASA, Norway. Printed 02.05. All rights reserved.
Dynal®, Dynabeads® and Dynal MPC® are registered trademarks of Dynal Biotech ASA.
The Dynabeads® 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: + 47 22 06 10 00
Dynal Biotech LLC, USA, tel: + 1 800 558 4511
Dynal Biotech GmbH, Germany, tel: + 49 40 36 15 73 23
Dynal Biotech Ltd, UK, tel: + 0 800 731 9037
Dynal Biotech S.A, France, tel: + 33 3 44 23 45 95
Dynal Biotech Pty Ltd, Australia, tel: + 1 800 623 453
Dynal Biotech Beijing Ltd, tel: + 86 10 67 80 67 08
Nihon Dynal K.K., Japan, tel: + 81 3 35 93 78 61
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MOL.B.017.01