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View additional product information for Clear Flat-Bottom Immuno Nonsterile 384-Well Plates - FAQs (464718)
10 product FAQs found
For energy treated plates, the entire plate surface receives the energy treatment. The MaxiSorp, PolySorp, MediSorp and MultiSorp plates are energy treated plates. For traditionally treated plates, the well surface is treated up to the listed working volume.
We have tested and found that a 3 amino acid peptide (Pro, Leu, Gly) cannot be detected when passively adsorbed on the MaxiSorp surface. However, this peptide can be detected when covalently immobilized using CovaLink NH Modules and CovaLink NH2 Modules and Plates. Using covalent immobilization of small peptide residues, you can typically obtain a better orientation of the molecule and reduced problems with antibody recognition of the peptide due to masking of the epitope. We have discovered that a 7 amino acid peptide from the MHC Class II antigen can be detected when adsorbed on the MaxiSorp surface. We state that the detection limitation using the MaxiSorp surface is between 3 and 7 amino acid residues.
One additional note is that detection is contingent upon the orientation of the peptide when immobilized. If the active site is inactivated or hidden at the site facing the solid phase, no detection signal is observed.
Single-stranded DNA can be adsorbed to our MaxiSorp surface using approximately 10 µg ssDNA per mL PBS, pH 8.2, although the stability is uncertain. Based on our experience, ssDNA immobilized on the MaxiSorp surface is so loosely bound that it can be removed by stringent washing.
Double-stranded DNA will not bind to the MaxiSorp surface. DNA, however, can be covalently bound to Nunc NucleoLink Strips.
Our Thermo Scientific 8-Well Strip Caps for Immuno Standard Modules were designed to provide a positive seal for flat and round bottom wells of our Nunc MicroWell 96-Well Microplates and Nunc Immuno 96-Well Plates.
8-Well Strip Caps for Immuno Standard Modules are not compatible with C or V bottom wells of the Nunc Microwell 96-Well Microplates and Nunc Immuno 96-Well Plates.
The following list describes the geometries of wells available for Nunc Immuno-plates and modules:
- Flat bottom (F): Allows maximum transmission of light. These plates can be read on a monochromatic reader.
- Round bottom (U): This geometry optimizes washing and coating. These plates can be read using a dual wavelength reader.
- "C" bottom (C): This design of the well is a combination of both flat and round bottoms. Basically, it is a flat bottomed well with curved edges at the bottom. These plates also can be read using a monochromatic reader combining optimal reading and washing.
- StarWell: These wells have a modified "C" shape geometry with eight fins strategically placed at the bottom. This feature increases surface area, allowing more molecules to become immobilized which reduces incubation times.
Both of these surfaces are identical. The only difference between them is that for the certified plates, a representative sample from each manufacturing lot undergoes a Binding Capacity test. This test is an ELISA-like assay used in our quality control laboratories to ensure binding capabilities.
Assay sensitivity depends strongly on efficient removal of non-specific reacting molecules. High background readings and coating instability can be minimized by addition of a blocking step after the first coating. The excess surface is then occupied by indifferent molecules.
We recommend washing three times after each coating step by using a solution of 0.15 M phosphate buffer (pH 7.2) with 0.2 M NaCl and 0.05% Tween 20.
For blocking, we recommend using 0.5% BSA, 1% casein or 1% gelatin in 0.15 M phosphate buffer (pH 8.2) or carbonate buffer (pH 9.6).
The choice of plates or modules will depend on the type of RIA you are planning to perform and specific assay conditions. Nunc provides two different surfaces for immunoassays, PolySorp and MaxiSorp.
- MaxiSorp surface is highly charged and should be used if the assay requires quantitative measurement of proteins (antibodies) or molecules with polar groups.
- PolySorp surface should be used if the assay consists of less polar molecules with hydrophobic characteristics.
Molecules bind to our PolySorp and MaxiSorp surfaces through passive adsorption.
Using IgG as a reference molecule and knowing that it is a globular molecule, theoretical calculations indicate that the maximum binding for proteins are:
- For the MaxiSorp surface, in monolayer: 650 ng/cm^2
- For the PolySorp surface: 220 ng/cm^2.
MaxiSorp, PolySorp and MiniSorp surfaces were developed for immunology assays:
- The MaxiSorp surface is a modified, highly charged polystyrene surface with high affinity for molecules with polar or hydrophilic groups. The surface has a high binding capacity for proteins, including globular antibodies in proper orientation. Thus, it offers very high sensitivity in double antibody "sandwich" tests.
- The PolySorp surface is more hydrophobic than the MaxiSorp surface. It has high affinity to molecules of a more hydrophobic character. This surface is particularly suited to non-protein antigens including virus antigens.
- The MiniSorp surface is a polyethylene surface with very low affinity to molecules of any type. This type of surface is ideal for the liquid phase immuno techniques.
- The MiniSorp surface is only available in a tube format, while the MaxiSorp and PolySorp surfaces are offered in both 96-well plate and module formats.