To begin, place a sealed bag containing frozen dried lectin fixed beads on a bench at room temperature. Place an antistatic cloth on the desk and then position the portable static eliminator facing the workspace. Using antistatic tweezers, insert the beads into an empty tip such that two spacer beads are placed in between the lectin fixed beads.
Use nippers to gently squeeze the top of the tip and close the opening. Manually straighten the tip if it is bent. Then store it in a sealed bag with a desiccant at four degrees Celsius until needed.
Add about 10 microliters of the analyte protein solution to a tube containing 10 micrograms of dried biotinylation reagent. Vortex the mixture to mix well. Then centrifuge it at 1000 G for five seconds at room temperature.
Incubate the tubes for one hour at room temperature away from light. Now add 90 microliters of probing buffer to the samples to dilute it tenfold before incubating. Open the front door of the automated reaction measurement device.
Transfer the liquid reservoirs, the prepared tips and the analytes into the designated positions in the device. Turn on the power switch of the main unit of an automated reaction measurement device. Touch the Start Diagnosis button displayed on the screen.
Press Assay on the home screen to select the protocol. Now select the BAssaySTD option of the assay method and press Next. Enter the sample names and press Next.
Then enter the tip names and press Next again. Confirm the notes on the right, then press Start to begin measurement. Confirm the screen switch to the running mode and wait until the Finalize arrow is highlighted.
The screen will change into result mode when the measurement is complete. Insert a USB memory stick into the port on the side of the main unit. Press Export in the result mode to save the data as a CSV file for further data analysis and individual graphing in Excel.
A distinct difference in the binding pattern of the lectins against the 12 polyacrylamide sugars was seen. Purified protein products that were subjected to the tip measurement showed increased reactivity of certain lectins to the saccharides.
