Surfaces Selection Guide

Type of Surface
and Code
Binding capacity Binding interaction Biomolecule properties Suggested use
Medium Binding

MB
100-200 ng IgG/cm2

A polystyrene surface less polar than the HB 8 one and with affinity to molecules of a more hydrophobic nature. The MB surface can better work with lipidic molecules

Large molecules (> 20 kD ) with large or abundant hydrophobic regions

Assays where the molecules that have to be coated present large hydrophobic regions or assays that need hydrophobic polipetides as coated molecule

High Binding

HB8
Up to 400-500 ng IgG/cm2

A polystyrene surface with high affinity to molecules with mixed hydrophilic/hydrophobic domains such as proteins and antibodies.The surface is optimized for antibodies (IgG)

Improves binding of medium to large molecules ( > 10 kD ) that are positively charged with or without hydrophobic regions

Assays where the adsorbed molecule must work in excess ( up to 400-500 ng/cm2) - e.g. ELISAs ( indirect ELISA test for IgG or ELISA capture test for IgM detection )
Very suitable to set up competitive tests ( molecule < 50 ng/cm2) -e.g. steroids/ hormones - for the ability to orientate the adsorbed molecule

NO Binding

NB
N.A.

None-Inhibits hydrophobic and ionic interactions

Significantly reduces protein binding

Certain procedures require a surface that is non binding. Many proteins,enzymes in particular may become activated or inactivated upon attachment to a surface. A non-binding surface can be ideal for assays requiring this feature


Coated Surfaces

Type of Surface
and Code
Binding capacity Binding interaction Biomolecule properties Suggested use
Biotin

Bio
N.A.

Streptavidin/biotin non covalent biological strongest interaction

Any streptavidin or avidin molecules

Interactions with avidin
interactions with streptavidin

Calmodulin

Cal
N.A.

Calmodulin binds proteins through interactions with its hydrophobic sites

Proteins involved in neurotransmission

interactions with proteins involved in glycogen metabolism
interactions with factors involved in neurotransmission mechanism
interactions with enzymes involved in the NAD*/NADP* phosphorilation system

Concanavalin A

Con A
N.A

Specific for C3-C4-C5 hydroxyl groups

Carbohydrates

High affinity for terminal a-D-mannosyl and a-D-glucosyl residues.
Used to immobilise glycoproteins or carbohydrates containing these groups

Heparin Catcher

HC
N.A.

A special surface onto which heparin can be immobilized

UHF heparin

The proposed heparin ELISA test is a quantitative enzyme-linked assay for the in vitro measurement of unfractionated heparin in low protein content fluid such as a buffer or urine

Jacalin

Jac
N.A.

High affinity for a-D-galactose residues of the biochemical structure of IgA1 and cell membranes

Human IgA 1

Human IgA1 specific binding, sterically oriented
Purification of human immunoglobulins (especially IgA1)
Separation of immunocomplexes antigen-antibody
Separation of IgA1 from contaminants Stimulation of T-cells

Neutravidin


4 biotin- binding sites with the highest specificity

Same of Streptavidin

Any biotinylated molecule

Same of streptavidin with the lowest non-specific binding

Poly-L-lysine
Poly-D-lysine
Lys-L
Lys-D
N.A.

This polymer creates a uniform net positive charge on the plastic surface

Cells and nucleic acids

This surface can enhance cell attachment, growth and differentiation.
Moreover this surface can tight bind negatively charged nucleic acids like dsDNA

Poly-L-Arginine

AR
N.A.

High density of: α-amino/α-carboxyl/ guanidino groups able to react trough electrostatic and stereospecific bonds

Affinity for prekallikrein,clostripain, prothrombin,plasminogen and plasminogen activator

interactions with serino proteases
interactions with maturation promoting factors

Protein A

PA
~ 5 pmol IgG/well

Specific for Fc region of IgG

Most immunoglobulins of mammalian species

- Binds strongly to IgG from human, rabbit,guinea pig,pig, dog and cat
- Binds strongly to mouse IgG2a,IgG2b and moderately to IgG3
- Binds to Fc region of antibodies for optimal orientation

Protein G

PG
~ 5.3 pmol IgG/well

Specific for Fc region of IgG

Most immunoglobulins of mammalian species

- Binds strongly to IgG from human, rabbit,mouse ,pig,bovine,dog , goat and horse
- Binds only to IgG- no cross-reactivity with other antibody classes
- Binds to Fc region of antibodies for optimal orientation

Streptavidin

SA 5/200
12 pmol of biotin/200 ul of coated volume The binding will vary depending on the nature of the biotin-bound moietes and the steric hindrance

Non covalent interaction towards biotinylated molecules; streptavidin-biotin interaction is nearly irreversible since the binding affinity ( Ka ) in solid-phase is around 108-1010

Any biotinylated molecule

Streptavidin coated microplate is designed for analysis of biotinylated molecules or for indirect coating with biotinylated ligands. This support binds very effectively any biotinylated molecule, with very rapid kinetic and excellent binding capacity, offering a multipurpose tool for various capture and immunodetection systems

Streptavidin High binding

SP75/100
From 15 to 40 pmol/100 ul of coated volume

Same as previous

Any biotinylated molecule

Competitive assays - when very small molecules must be captured

Wheat germ

WG
N.A.

Specific binding of N-acetyl-D-glucosamine residue

Glicoproteins, enzymes and cell membranes

Studies of surfaces of normal and transformed cells
Glycoprotein purification including membrane glycoproteins
Studies of cell surface changes during development and the cell cycle

Maleimide

MAL
free sulfhydryl groups (e.g. Peptides that contain a terminal cysteine or thiol containing haptens) , or reducible disulfide bonds 

offer a powerful instrument for binding biomolecules containing free sulfhydryl groups (e.g. Peptides that contain a terminal cysteine or thiol containing haptens) , or reducible disulfide bonds that are difficult to coat onto polystyrene plates 

 

assays requiring site-directed orientation of particular biomolecules especially during antibody production.


Chemically Activated Surfaces

Type of Surface
and Code
Binding interaction Biomolecule properties Suggested use
Aminated with primary and secondary amines
AM 1 / AM2

Reactive amino groups allow immobilization of molecules through reductive amination of aldeyde or ketone groups without prior activation of the surface.Alternatevely, molecules can be immobilized through amide-bond formation with carbodiimide-activated carboxyl groups. Bifunctional crosslinkers may be used to introduce other functional groups.

Proteins,peptides, carbohydrates, enzymes

Immobilisation of molecules which are bound weakly or not at all by physical adsorption, namely small peptides (M.W. 1000-5000) drugs, toxins or hormones.
Oriented immobilisation of molecules in order to secure the integrity and accessibility of their specific sites avoiding the risk of inhibition of these sites by casual physical adsorption for such molecules as Fab-SH-antibody fragments, or nucleic acids (single or double strand)

Carboxylated
COOH

Carboxyl groups can be activated with a carbodiimide followed by coupling of an amino containg molecule, resulting in a stable amide bond between the surface and the molecule. Alternatevely bifunctional crosslinkers may be used to introduce other functional groups.

Molecules containing amino groups

The amino group presents in any molecules, such as peptides or proteins, binds to COOH surface through formation of amide bonds between the amino group presents in the molecule and the surface carboxylic group by the action of carbodiimide .