Concanavalin A Slide Coating Protocol

 

Introduction

We initially developed a slide coating method using Concanavlin A, a prototypical lectin that binds glycosul and mannosyl groups. Although we favor custom made slides from Erie Scientific for our arrays, home made slides can be easily made for small scale experiments.

Materials for Microarray Printing
Item
Product Number, Supplier
Use
Concanavalin A
L7647, Sigma
For "home-made" Concanavalin A coated slides
VWR Slides
48300-025, VWR
For "home-made" Concanavalin A coated slides
Ambion Nuclease Free Water
9332, Ambion
For "home-made" Concanavalin A coated slides

Protocol

Concanavalin A Solution

  1. Measure 0.0625 g of Concanavalin A (ConA).
  2. In a graduated cylinder, add ConA to 200 mL nuclease free water. Mix by inversion.
  3. After ConA is dissolved, bring volume to 250 mL.
  4. Pour ConA into a 250 mL jar. Add a sterilized stir bar to jar, seal top of jar and stir ConA on a stir plate for 4 hours at a speed that creates a sufficient vortex in the solution.
  5. After 4 hours of stirring, filter solution into a new, sterilized container with 0.22 um vacuum filter.

Slide Coating (Performed in tissue culture hood. Use powder free gloves when manually handling slides)

  1. In a tissue culture hood, pour filtered ConA solution into a slide dipping tub that can accomodate 10 slides.
  2. Spray the fingertips of the gloves used to handle the slides with 100% ethanol.
  3. Load 10 pre-cleaned glass slides into dipping cartridge.
  4. Dip slides up and down in the ConA solution 5 times (1 second each dip), then submerge slides in ConA for 2 minutes.
  5. Remove slides from dipping cartridge and lean slides (vertically) against a pipette tip box in the tissue culture hood.
  6. Allow slides to dry in this fashion for 1 hour.
  7. Fill 2 dipping tubs with nuclease free water (Tissue Culture grade water can be used here). Dip slides (up and down for 1 second) 5x in first tub, then dip 1x in second tub.
  8. Dry slides for 1 hour as before.

Slide Storage

  1. Store slides in sterilized slide boxes in a vacuum dessicated container.
  2. Slide quality is optimal for 1 week after production.