Procedure for Coating Microarray Slides with MCP-2, MCP-4, or MCP-2F
Download the MCP2/4 Guide here
This guide is written for glass microarray slides. Many surfaces with free hydroxyl groups can be coated similarly. For surfaces naturally lacking hydroxyls, they must first be introduced by treatment as described below.
Chemical pre-treatment:
Treating the substrate, before coating, is recommended.
If available, treat with oxygen plasma for 10 min. Otherwise, follow these instructions.
Submerge slides in 1.0N NaOH (1 hr.)
Rinse with water.
Submerge in 0.1N HCl (20 min.)
Rinse with deionized water.
Instructions for coating slides with MCP-2/2F/4 copolymer
-
First, dilute the provided MCP copolymer with the proper volume of Coating Solution (#5XCOT1G) in a glass beaker. Stir well, or vortex, to mix. Prepare this solution immediately prior to use.
Use the following ratios based on your copolymer product:
1 mL MCP-2 20 mL 5XCOT1G with 30 mL filtered water 1 mL MCP-4 10 mL 5XCOT1G with 40 mL filtered water 1 mL MCP-2F 6 mL 5XCOT1G with 44 mL filtered water Note: Some cloudiness may occur when diluting MCP solution for coating. This does not appear to affect the performance of the coating.
Second, immerse the slides in the copolymer solution for 30 min. at room temperature.
Third, wash slides individually in a large volume of water. For small numbers of slides, grasp each by forceps and swirl for a few seconds in 1 L deionized water.
Finally, and immediately, dry the slides. You may use a stream of nitrogen, or in a slide-rack centrifuge to remove water droplets and spots. It is very important to get rid of spots and droplets when drying, to avoid irregularities.
To complete drying, dry the slides at 80°C in a vacuum oven (<2 mm Hg) for 15 min.
Instructions for Storage of coated slides
Store inside a vacuum-sealed bag, with a desiccant pack or in a desiccator. Store frozen (-20°C or lower). Under these conditions, coated slides are stable for at least 1 year.
Suggested Spotting Guidelines
Control relative humidity (rh). Lower rh (30-45%) works best. A tray of desiccant inside the arrayer can help control rh on humid days.
Bake substrates at 80°C 15 min. immediately before spotting.
-
50 mM trehalose in spotting buffer may slightly increase spot diameter, but leads to greater uniformity within spots.
An optimized Spotting Buffer is available from Lucidant (#SPT1). This is generally used for oligos, although it may improve spotting with some proteins as well. For most proteins, phosphate buffer works well.
Suggested Blocking Guidelines
After spotting, block remaining reactive groups with Blocking Solution (Lucidant #BLK1) 30 min. at room temperature (50°C for oligonucleotide arrays).
Rinse well with deionized water.