Wednesday, September 14, 2011

Microarray Printing Concentration

A simple test to determine the best concentration of protein to print onto SuperNHS Microarray Substrates:  Whole Human IgG is printed in Arrayit Protein Printing Buffer using 946MP3 Microarray Printing Pins and a NanoPrint Microarrayer.  Three different print runs left to right top down with final concentrations in triplicate at 0.5, 0.25, 0.125, 0.0625, 0.03125 µg/µl final concentration.  16-bit raw data above presented in pseudo color rainbow pallet provided by ImageJ. Microarrays were processed in an AHC4x24 and reacted with 1/1000 dilutoin of anti-human IgG conjugated to a fluroescent dye and scanned with an Innoscan 710AL Microarray Scanner, gain 1, laser power low, at 10 micron resolution. 0.5 µg/µl concentration protein works best to fully saturate the binding sites of the surface chemistry as that provided saturated signals at the lowest possible sensitivity settings on the scanner. To download and view the consistant raw data from all 96 microarrays in this test point your browser to: http://www.arrayit.com/Arrayit_Corporation/SuperNHS-data.zip