Surface Activated Dynabeads.indd - Invitrogen

Surface Activated
Dynabeads ®
www.dynalbiotech.com

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A variety of ways to bind your ligand
Manual or automated handling
Protein separation technology
Introduction
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).
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.
Hydrophobic or
Hydrophilic
Dynal Biotech provides surface
activated Dynabeads ® 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.
Fig. 1 Principle of biomagnetic
separation with Dynabeads ® .
Surface Activated Dynabeads ®
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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.
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 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 Dynabeads ® are easy to
handle manually using simple magnet racks for the isolation
of target.
Dynabeads ® 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.
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Ligand Target
Antibody Protein or
antibody
Antibody
fragment
Low MW
antigen 1
or peptide
Dynabeads ®
Tosylactivated
Dynabeads ®
Carboxylic
Acid
Dynabeads ®
Amine
Dynabeads ®
Epoxy
√ √ √ √ √ √ √
√ √ √ √ √ √ √ √ √
Dynabeads ®
Subcellular
Organelles √ √ √ √ 2 √ √ √
Bacteria √ √ √
Virus √ √ √ √ √ √ √ √ √
Phage 1 √ √ √ √ √ √ √ √ √
Phage 1 or
antibody
Protein Phage 1 or
carbohydrate
Peptide Phage 1 or
antibody
√ √ √ √ √ √ √ √ √ √
√ √ √ √ √ √ √ √ √
Nucleic acid √ √ √ √ √ √
√ √ √ √ √ √ √ √ √ √
Carbohydrate Antibody √ √ √ √ √
Low MV antigen Antibody √ √ √ √ √ √ √ √
Nucleic acids,
oligonucleotide,
aptamer, PNA
Nucleic acid
binding
proteins 1
DNA 1 , RNA 1 or
PCR amplicants 1
Enzyme Substrate
or target
for enzyme
degradation
Organic
chemistry
derivatisation,
including
introduction of
new functional
groups
1 Dynabeads ® Streptavidin may be the best product choice
2 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
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√ √ √ √ √ √ √
√ √ √ √ √ √ √ √
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Surface Activated Dynabeads ® : Dynabeads ® M-500
Subcellular
Bead Characteristics: • Slightly hydrophobic bead.
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Surface tosyl groups.
Bead diameter 4.5 µm.
Binding Properties: • Direct covalent binding to
primary amino- or sulphydryl
groups in proteins and
peptides.
No further surface
activation required.
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.
Applications and
Reaction Chemistries:
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The Fc portion is generally
more hydrophobic than
the Fab regions, facilitating
biologically active orientation
of your antibody.
Organelle fractionation.
EM pictures of organelles.
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Dynabeads ® M-280
Tosylactivated
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Hydrophobic bead.
Surface tosyl groups.
Bead diameter 2.8 µm.
Direct covalent binding to
primary amino- or sulphydryl
groups in proteins and
peptides.
No further surface
activation required.
Binding overnight at neutral
to high pH and 37 o C.
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.
Protein purification.
Isolation of fragile cells
(the smaller beads are
pulled gently to the magnet,
ensuring the isolation of
intact and viable cells).
IVD
Dynabeads ® MyOne TM
Tosylactivated
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Hydrophobic bead.
Surface tosyl groups.
Bead diameter 1.0 µm.
Direct covalent binding to
primary amino- or sulphydryl
groups in proteins and
peptides.
No further surface
activation required.
Binding overnight at neutral
to high pH and 37 o C.
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.
Protein purification.
Isolation of rare cells (rapid
kinetics due to high number
of beads).
IVD
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Dynabeads ® M-270
Amine
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Hydrophilic bead.
Surface amino groups.
Bead diameter 2.8 µm.
Direct covalent binding
through reductive amination
of aldehydes.
No further surface
activation required.
Rapid binding (less than 1
hour) at neutral to high pH
and room temperature.
Rapid immobilisation of
carbohydrates, glycoproteins
and glycolipids
(e.g. lipopolysaccharides).
Easy introduction of
further alternative surface
chemistries.
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
peptides.
Dynabeads ® M-270
Carboxylic Acid
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Hydrophilic bead.
Surface carboxylic acid
groups.
Bead diameter 2.8 µm.
Covalent amide bond
formation with primary
amino- and sulphydryl groups
in proteins and peptides.
Activation through
carbodiimide is required.
Immediate peptide bond
formation at pH 5-6 and
room temperature.
100% covalent binding.
Low non-specific binding of
nucleic acids.
Rapid binding chemistry.
Immobilisation of labile
proteins and peptides.
N-terminal coupling of
peptides.
IVD
Dynabeads ® MyOne TM
Carboxylic Acid
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Hydrophilic bead.
Surface carboxylic acid
groups.
Bead diameter 1.0 µm.
Covalent amide bond
formation with primary
amino- and sulphydryl groups
in proteins and peptides.
Activation through
carbodiimide is required.
Immediate peptide bond
formation at pH 5-6 and
room temperature.
100% covalent binding.
Low non-specific binding of
nucleic acids.
Rapid binding chemistry.
Immobilisation of labile
proteins and peptides.
N-terminal coupling of
peptides.
IVD
NA IVD
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Dynabeads ® M-270
Epoxy
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Hydrophilic bead.
Surface epoxy groups.
Bead diameter 2.8 µm.
Direct covalent binding to
primary amino and
sulphydryl functional groups
in proteins and peptides.
No further surface
activation required.
Binding overnight at neutral
pH, high salt and a wide
temperature range.
The functionality of your
enzyme is maintained
after isolation.
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.
www.dynalbiotech.com

Ordering Information
Product Concentration Volume Product No.
Dynabeads ® M-270 Amine
Hydrophilic beads with amino groups.
Dynabeads ® M-450 Epoxy
Hydrophobic beads with epoxy goups.
Dynabeads ® M-270 Epoxy
Hydrophilic beads with epoxy goups.
Dynabeads ® M-450 Tosylactivated
Hydrophobic beads with tosyl group.
Dynabeads ® M-280 Tosylactivated
Hydrophobic beads with tosyl group.
Dynabeads ® MyOne Tosylactivated
Hydrophobic beads with tosyl group.
Dynabeads ® M-270 Carboxylic Acid
Hydrophilic beads with carboxylic acid groups.
Dynabeads ® MyOne Carboxylic Acid
Hydrophilic beads with carboxylic acid groups.
Dynabeads ® M-500 Subcellular
Slightly hydrophobic beads with tosyl groups.
© 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.
30 mg/ml
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 www.dynalbiotech.com
www.dynalbiotech.com
MOL.B.017.01
2 ml
10 ml
143.07
143.08
30 mg/ml 5 ml 140.11
60 mg
300 mg
freeze-dried
freeze-dried
143.01
143.02
30 mg/ml 10 ml 302.12
30 mg/ml
30 mg/ml
100 mg/ml
100 mg/ml
30 mg/ml
10 mg/ml
2 ml
10 ml
10 ml
2 ml
10 ml
2 ml
10 ml
2 ml
10 ml
100 ml
142.03
142.04
301.01
655.01
655.02
143.05
143.06
650.11
650.12
650.13
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.