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Functionalized 96-well Plates For Microarrays

PolyAn offers surface modified 96-well plate products that have been equipped with PolyAn’s 3D-Epoxy and 3D-NHS matrices, respectively, for covalent coupling of DNA, peptides and proteins:

ID Surface Modifications Format
00 680 251 3D-Epoxy 96 well plate, white, 12 x 8-strip, flat bottom
00 680 451 3D-NHS 96 well plate, white, 12 x 8-strip, flat bottom
00 690 251 3D-Epoxy 96 well plate, transparent, 12 x 8-strip, flat bottom
00 690 451 3D-NHS 96 well plate, transparent, 12 x 8-strip, flat bottom (i)
00 691 251 3D-Epoxy 96-Well µClear® Microplatte, PS, schwarz
00 692 251 Covalently coated Streptavidin 96 well plate, transparent, 12 x 8-strip, flat bottom
00 693 251 Covalently coated Neutravidin 96 well plate, transparent, 12 x 8-strip, flat bottom

The three-dimensional functional matrix allows the development of covalent binding of nucleophiles to the plate surface without complex coupling chemistry. The structure of the matrix, which has a thickness of up to 50 nm, also reduces unspecific binding to a minimum.

3D-Epoxy surface chemistry
for coupling via nucleophilic groups (e.g. N-terminus) of biochemical species


The 3D-Epoxy 96 well microplates are used mainly if adsorptive binding of peptides or oligonucleotides, for example, to high/medium binding surfaces is ineffective or the binding strength is not sufficient. Areas of application include detection methods such as ELISA, ELI-Spot, protein and peptide arrays and DNA binding.

A covalent bond is formed by sharing of electrons between two atoms. The dissociation energy for a typical covalent bond is 100 kcal/mol and by far the strongest in chemistry.

Epoxides are cyclic ethers with a highly strained three member ring. Epoxy rings can be easily reacted with nucleophiles e.g. amines, hydrazines, thiols, hydroxides and carboxyl groups. Compared to NHS-esters or 1,4-Phenylene isothiocyanates (PDITC) the epoxy surface is more stable and has a longer shelf-life. Epoxy-surfaces are stable up temperatures of 40°C and are also more stable against humidity compared to NHS and PDITC-surfaces.

The nucleophilic addition is catalyzed by acid or basic conditions. Under acidic conditions, the oxygen in the ring is positively charged, which facilitates the nucleophilic attack. Under basic conditions the least substituted carbon is attacked by the applied nucleophile in a standard SN2 reaction.

3D-NHS surface chemistry
for coupling via the N-terminus of biochemical species


The NHS-ester reacts immediately with the NH2- terminus of biochemical species to form a covalent bond with the surface (420 kJ/mol). The reaction of carboxyl functionalities with N- Hydroxy succinimide leads to highly reactive esters, which can be easily reacted with nucleophiles e.g. amines, hydrazines. However, due to its high reactivity the NHS ester is susceptible against hydrolysis and is characterized by a relatively short shelf-life. All NHS-activated surfaces should therefore be processed quickly.

There are a number of different approaches to couple on the NHS-surface:

  1. It is assumed that not all Carboxy groups have reacted to NHS- esters during activation. Thus a negatively charged carboxy surface still remains which in turn supports the physico-chemical adsorption of positively charged probes e.g. NH3+. Hence a protonating media (pH < 5) for the biochemical species getting a positively charge is required.
  2. A nucleophilic attack on the active ester is also catalyzed under basic conditions (pH > 8,5).

After attachment of the biochemical species the surfaces must be blocked with a blocking buffer containing small molecules that can access all reactive groups within the 3D-Matrix.

Custom Product Request

Contact us if you require a special surface for binding of your biomolecules that is not listed in the products table. We can also functionalize other plate formats and substrates with our surfaces.

Additionally, we have access to a wide range of different surface modifications for binding of small molecules, saccharides etc.
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