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What Is A 96 Well Plates?

Posted on: August 10, 2020, by Jason

96 Well Plates

The 96 well plates is one type of ELISA PLATE, is a key role in the Enzyme Linked Immunosorbent Assay (ELISA). It's involved in the immunological reaction, such as: antibody or antigen coating, buffer block, etc.

The well plate is manufactured in a GMP clean room. The product quality is strictly controlled in accordance with the requirements of ISO and ANSI/SLAS Standards to ensure the stability of the product batches. The product variation coefficient of each plate hole and batch is less than 5%. The product adopts optimized surface treatment technology, which effectively improves the adsorption capacity of the protein in the high and medium binding capacity ELISA plate, and improves the sensitivity of the product to protein adsorption detection. It can be widely used in various types of ELISA, PCR detection, Cell culture, scientific research, etc.

Types Of 96 Well Plates

According to the microplate reader, it is divided into detachable and non-detachable.

Non-Detachable 96 well plates

For the non-detachable, the well strips on a whole board are connected together.

Detachable 96 well plates

Then the detachable is that the well strips on the board are separated, and the separated strips are 8 wells or 12 wells per strip.

Generally, detachable microplates are commonly used. Although all well plats has same apperance, but different brands have their own technical difference, such as structure, protein adsorption performance, etc. While you choose to buy 96 well plates, you should also consider which brand of microplate reader is better you.

According to the different binding ability with protein and other molecules, the ELISA plate can be divided into the following categories:

High Binding Well Plates

After surface treatment, the protein binding capacity of the ELISA plate is greatly increased, reaching 300-400 ng 1gG/cm2, and the molecular weight of the main binding protein is >10kD. To  use this type of well plates can improve the sensitivity, and can relatively reduce the concentration and dosage of the coating protein. The disadvantage is that it is easier to produce non-specific reactions. After the antigen or antibody is coated, non-ionic detergents cannot effectively seal the unbound protein sites, and protein should be used as a sealant.

Medium Binding Well Plates

The ELISA plate is passively combined with white through the hydrophobic bond on the surface, and is suitable as a solid-phase carrier for macromolecular proteins with a molecular weight of 20D. Its protein binding capacity is 200-300 ng 1gG/cm2. Due to the characteristic of ELISA plate that only binds to macromolecules, it is suitable as a solid-phase carrier for unpurified antibodies or antigens, and can reduce potential non-specific cross-reactions. This microplate can be used as a blocking solution with inert protein or non-ionic detergent.

Animated Enzyme Well Plates

After surface modification, the alcohol standard has a positively charged amino group, and its hydrophobic bond is replaced by a water-killing bond. It is suitable as a solid phase carrier for small molecule proteins. Using a suitable buffer and pH value, the surface can bind to small negatively charged molecules through ionic bonds. Due to the hydrophilic properties of the surface and the ability to covalently bond with other crosslinking agents, it can be used to immobilize protein molecules soluble in detergents such as Triton-100 and Tween20. Its disadvantage is that due to the reduced hydrophobicity, some protein molecules cannot bind; in addition, its surface needs to be effectively sealed. Due to the hydrophilic and covalent surface characteristics, the sealing fluid used must be able to interact with the non-reactive amino groups and any functional groups in the selected crosslinking agent.

How to detect the protein adsorption of the microplate?

A protein solution of known concentration can be used for coating on a blank well plate for ELISA. Measure the protein concentration of the solution in the well after coating. The amount of protein in the well before coating minus the amount of protein after coating is the amount of protein adsorbed on the plate. In this way, we can know the adsorptivity of the microplate.

Raw Materials Of 96 Well Plates

There are 4 types materials: Polystyrene, PVC, Poly-D-Lysine and Quartz. Polystyrene is commonly used to make 96-well plates, but polystyrene has poor chemical stability and can be dissolved by a variety of organic solvents (such as aromatic hydrocarbons, halogenated hydrocarbons, etc.), and will be corroded by strong acids, alkalis and anti-grease. easy to change color after being irradiated by ultraviolet light, so be careful during use.

The Apperance Of 96 Well Plates

Wells: 96 wells

well is the most commonly used now, because the microplate is used with microplate reader.

Struture: Detachable and non-detachable (SOLID)

elisa-plate
elisa-plate

Colors:  Clear, White, Black

micro-plate
micro-plate

Botton:

  • F-Botton (most commonly used);
  • U-Botton (for good mixing),
  • V-Botton (for microtiter use),
  • C-Botton (flat center with rounded corners to improve fluid recovery)
microplates
microplates

The Components of 96 Well Plates

The Board (3 colors: clear, white, black)

microplates-96-well-plates
microplates-96-well-plates

Well Strip (3 colors: clear, white, black)

well-plates-well-strip
well-plates-well-strip

8 well breakable strip and 12 well breakable strip

The Lid

well-plates-lid
well-plates-lid

The specification of 96 well plates

96 well plates
96 well plates

405nm absorbance test result of blank 96 well plates

Well plates

How much can a 96 well plates hold?

  • Well Plate diameter: 6.8mm;
  • 96 well volume: 360 μL;
  • Working volume: 75-250 μL;
  • Max. volume: 24,000 μL per well plate

96 Well Plate Dimension

96-well-plates-dimension
96-well-plates-dimension

According to the ANSI/SLAS-Standards 1 to 4-2004, the standard 96 well plates dimension are 127.76mm(L) x 85.48mm(W). It's internationally recognized standards, includes 5 standards as below:

ANSI SLAS 1-2004 (R2012): Footprint Dimensions
last updated January 9, 2004

ANSI SLAS 2-2004 (R2012): Height Dimensions
last updated January 9, 2004

ANSI SLAS 3-2004 (R2012): Bottom Outside Flange Dimensions
last updated January 9, 2004

ANSI SLAS 4-2004 (R2012): Well Positions
last updated January 9, 2004

ANSI SLAS 6-2012: Well Bottom Elevation
last updated April 09, 2009

What are 96 well platess used for?

1. The solid-phase polystyrene surface as a carrier also plays an important role in the adsorption of antigens, antibodies or antigen-antibody complexes.

2. Antigens, antibodies and other biomolecules are adsorbed to the surface of the carrier through a variety of mechanisms, including passive adsorption through hydrophobic bonds, water/ionic bonds, covalent bonding through the introduction of other active groups such as amino and carbon groups, and through Hydrophilic bonding after surface modification.

Used in various experimental platforms: IVD reagents, Lateral Flow Assay, ELISA, PCR, nucleic acid detection, DNA extraction, cell culture, scientific research, etc.

How are 96 well platess used in food safety testing?

It’s widely used in food contamination with ELISA test, PCR test and Lateral Flow Test. The 96 well plates popular used in milk test kit in raw milk detection antibiotic residues. Unlike the traditional lateral flow design, the labled Colloidal Gold will not dispens on the nitrocellulose membrane, it’s separated from strip, load on the 96 well plates help increase sensitivity, fast dilute analyte and shorten the detection time.

96-well-plate-elisa
96-well-plate-elisa

The famous brand of 96 well plates

  • BRAND MICROPLATE
  • CORNING MICROPLATE
  • NUNC MICROPLATE
  • GREINER MICROPLATE
  • COSTAR MICROPLATE

Conclusion       

Well plates is a commonly laboratory consumables, except for 96 well plates, there are some types of microplates: 6 well plates, 12 well plates, 24 well plates, 48 well plates, 96 well plates, 384 well plates, 1536 well plates. 

well-plates
well-plates

Reference

Microplate Standards
Wikipedia: Microplate
Microplate PDF

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