Thermo Scientific™ Black 96-Well Immuno Plates

Catalog No.	Surface Treatment	Volume (Metric) Working
12-566-24	MaxiSorp	350 μL
12-566-30	PolySorp	350 μL
14-245-197A	Medium Binding	280 μL
14-245-177	High Binding	280 μL
21-377-271	Universal Binding	350 μL

Catalog No. 12-566-24 Supplier Thermo Scientific™ Supplier No. 437111

Description

Perform quantitative and qualitative immunoassays, like ELISA and binding assays, with these black, 96-well immuno plates, available in a choice of three surface types.

Perform quantitative and qualitative immunoassays, like ELISA and binding assays, with Thermo Scientific™ Black 96-Well Immuno Plates. Featuring a choice of three surface types and optimal binding, these plates minimize background in fluorescence reading.

Features:

• Plate has SBS footprint — fits standard equipment
• Flat-bottom wells
• Certified binding homogeneity/reproducibility

Choice of surface:

• Thermo Scientific PolySorp, Microfluor 1, Microlite 1+ and Universal Binding (UB) are for absorption of hydrophobic molecules.
• Thermo Scientific MaxiSorp is hydrophilic and ideal for antibody sandwich assays.
• Microfluor 2 and Microlite 2+ are slightly hydrophilic and bind a diverse range of biomolecules.

Compatible with:

Compatible with lids for Thermo Scientific MicroWell plates, sealing tape and breathable membrane

Ordering Information:

Availability may vary by country

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Specifications

• Binding Property: Hydrophilic
• Format: Standard
• Lid: Without Lid
• Detection Method: Fluorescence
• Array: 8 x 12
• Packaging: 80 Cases
• Binding Type: Adsorption
• Color: Black
• Product Type: Microplate
• Material: Polystyrene
• No. of Wells: 96
• Volume (Metric) Working: 350 μL
• Surface Treatment: MaxiSorp
• Well Shape: Flat Bottom (F-well)
• For Use With (Application): Fluorescence
• No. per Case: 80
• No. per Pack: 10
• Plate Blocking: None
• Target Molecule: Antibodies (Glycoproteins)
• Well Design: F96

Frequently Asked Questions (FAQs)

For Clear Flat-Bottom Immuno Nonsterile 96-Well Plates, is the surface treatment solely on the bottom of the well, or is it also applied to the sides?

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.

What length of peptide is ideal for binding to the Thermo Scientific MaxiSorp surface? What are the detection limitations?

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.

Can I bind either single- or double-stranded DNA to the Thermo Scientific MaxiSorp surface?

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.

Are your Thermo Scientific 8-Well Strip Caps for Immuno Standard Modules compatible with all Nunc MicroWell 96-Well Microplates and Nunc Immuno 96-Well Plates?

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.

For Nunc Immuno plates and modules, what are the advantages of one well geometry type over another? Which types should I use for various applications?

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.

What is the difference between certified and non-certified MaxiSorp plates and modules?

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.

How can I reduce high background readings and non-specific binding, when performing ELISA using Nunc-Immuno MaxiSorp Plates?

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).

Which surfaces do you recommend for RIA (radioimmunoassay)?

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.

Which 96-well Immuno plates or modules do you recommend for various applications?

The following list offers a brief description of the features of Nunc products and their specific applications:
- Nunc-Immuno Plates, MaxiSorp surface - These plates are designed for solid phase immuno assays and have a polystyrene surface with high affinity for polar groups and hydrophilic molecules. These 96-well plates are available with flat (F), round (U), or (C) bottom-well designs.
- Nunc-Immuno Plates, PolySorp surface - These plates have a polystyrene surface which adsorps less polar molecules compared to the MaxiSorp surface and has a high affinity for hydrophobic groups. These 96-well plates are available with flat (F), round (U), or (C) bottom-well designs.
- Nunc-Immuno Modules - These modules are designed for solid-phase immuno assays. The modules are available in 8-, 12- or 16-well formats with (F), (U), and (C) bottom-well and 8-well BreakApart with (C) bottom-well designs. These different formats allow one to choose the style which is appropriate for their assay design.
BreakApart Modules and LockWell Modules can be used for radioimmunoassays.
- Nunc StarWell Immuno Breakable Modules - The 8-well modules feature eight fins on the inner wall of the (C) bottom wells. This design increases the surface area by 50%. The increase in surface area allows more molecules to be immobilized, increasing assay signal. The fin configuration provides shorter diffusion distance to the surface, reducing incubation times.
- LockWell Immuno Breakable Modules - Each plate consists of 1 x 8 breakable strips. These strips are assembled in a designed frame which locks each well into place by a spring lock. This spring lock design orientates each well at the same horizontal level allowing uniform washing and reading. The LockWell Modules are available with round (U), (C) or StarWell bottom-well designs.
- Plates and Modules with (CovaLink) Covalent Binding Surfaces: These plates can be used to bind proteins, peptides, DNA or carbohydrates. Covalent linkage occurs via secondary amine group. This allows a specific orientation and provides improved stability compared to passive adsorption due to chemical binding between molecules. Binding of molecules on PolySorp and MaxiSorp surfaces is obtained by passive adsorption. CovaLink is a surface grafted with secondary amino groups (<NH) which serves as a bridge for covalent coupling. The covalent binding can immobilize small molecules such as biotin or peptides which may otherwise bind weakly by physical adsorption and facilitate the recognition by detection molecule.
- Nunc FluoroNunc/LumiNunc 96-Well Plates: These plates are optimized for IFMA (Immunofluorometric Assays) or FIA (Fluorometric Immuno Assays). The transparent polystyrene plates give a low background fluorescence and are optimal where a read-through system is used. The white plates/modules provide maximum reflection of fluorescence signal while maintaining low background. The white plates/modules are often used for epifluorescence reading. Black modules reduce background fluorescence and minimize back-scatter light which is often encountered in epifluorescence.

What is the maximum binding capacity for proteins on the Thermo Scientific MaxiSorp surface and PolySorp surface?

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.

What is the difference between your Thermo Scientific PolySorp, MaxiSorp, and MiniSorp surfaces?

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.

Should I use an opaque black/white immuno plate, or an "optical-bottom" design?

Opaque plates can only be used on “top reader” instruments, which detect (or, in fluorescence mode, excite and detect) from above the plate. Readers that detect from underneath the plate (or imaging systems) use optical-bottom plates, which have a black or white upper well structure and a transparent (polystyrene film or optical coverglass) bottom.

Why would I use a black or white (instead of clear) immuno plate?

Black plates are often used in assays that make use of a fluorescent reporter because the opaque black surface effectively isolates the signal from each well in order to eliminate “crosstalk” or signal bleed. White plates also isolate well-to-well signals somewhat, but also provide a reflective background to direct luminescent output to the reader, and so often serve as a better choice to improve dynamic range from a luminescent reporter.