UltraCoat II™ ELISA Plate Coating Buffer 5X

Store at 2-8°C. The solution may precipitate when stored at this temperature; if this occurs gently warm or store at room temperature until all crystals are dissolved.

Next Day 2-8°C

Stable for up to 6 months after dilution and for up to 18 months prior to dilution when properly stored at 2-8°C.

UltraCoat II™ ELISA Plate Coating Buffer is an enhanced coating buffer formulation designed for optimal coating of antibodies and antigens to polystyrene plates. Antibodies and antigens coated to polystyrene plates are commonly denatured or rendered non-functional and non-recognizable during plate drying. UltraCoat II™ ELISA Plate Coating Buffer helps stabilize immobilized proteins to preserve three dimensional structure and functionality.

• Increased Specific Signal and Decreased Background – UltraCoat IITM ELISA Plate Coating Buffer has an optimal pH and ionic strength designed for increased adsorption and consistent plate coating for the highest specific binding and reduced non-specific interference.
• Improved Assay Reproducibility – Coat plates at the same time and depending upon the activity of the coated protein, use one batch of coated plates for several months or years when stored dry at 2° - 8°C.
• Save Valuable Coating Protein and Detection Conjugate – By stabilizing coated proteins and preserving their tertiary structures, the specific signal will be increased and you may find that you can use less protein to coat your plates and less detection conjugate to obtain optimal signal.
• Ease of Use – Simply add 1 part UltraCoat IITM ELISA Plate Coating Buffer to 4 parts high purity deionized water. Following dilution, the buffer is ready to use and antimicrobial agents will suppress bacterial growth in the solution for 1 month at room temperature and for 6 months at 2° - 8°C.

1. Prior to use, simply add 1 part UltraCoat II TM to 4 parts high purity deionized water and mix for 15 minutes.
2. Add your antibody or antigen into enough coating buffer to coat the desired number of microwells (coating volume can range from 50 – 300 µl/well). Appropriate coating concentrations will range from < 0.1 µg/ml to > 10 µg/ml and each investigator should determine their own optimal coating concentration for their specific application. Stir this solution for 10 – 15 minutes before coating.
3. Pipette the desired coating volume into the microwells (50 – 300 µl/well).
4. Allow plates to incubate with coating solution for 3 – 24 hours at room temperature (25°C) protected from light.
Wrap the plate(s) in plastic wrap or incubate in a covered humidified chamber to minimize evaporation. 5. Dump or aspirate the coating solution out of the wells and wash plates > 4 times to minimize background staining.
6. Pipette an appropriate blocking buffer, such as UltraBlock-FISH™ ELISA Blocking Buffer “Ready Use” ( Leinco Prod. No.: B396), onto the plate at a higher volume than the coating solution (200 – 300 µl/well).
7. Allow plates to incubate with blocking buffer for 3 – 24 hours at room temperature (25°C) protected from light. Wrap the plate(s) in plastic wrap or incubate in a covered humidified chamber to minimize evaporation.
8. Aspirate the blocking solution from the wells.
9. The assay can be performed at this point or the plate(s) can be dried and sealed for later use.
10. Dry the plate(s) loosely covered with aluminum foil on the bench top or in a drying oven at room temperature or warmer for 2 – 24 hours.
11. Once plate(s) are dry, seal in an air-tight foil pouch with a desiccant and store at 2° - 8°C.