Alexafluor 647-labeled Streptavidin was used to detect biotinylated anti-mouse secondary antibody.
Hybridization protocol
Antibodies were incubated with the peptide microarrays for 1 hour at room temperature according to Stafford et al. (Stafford, Halperin, Legutki, Magee, Galgiani, Johnston, Physical characterization of the Immunosignaturing Effect, MCP 11(4), 2012).
Scan protocol
Scanning was done on an Innopsys Innoscan 910 at 10um resolution, high laser power, 70% PMT, autofocus ON, 545nm.
Description
Alexa-647 conjugated Streptavidin tertiary P. mallei_1
Data processing
Median normalization per slide was done, and any negative values were adjusted to 0.01.
In situ synthesis of peptide microarrays - shadow mask design
Data table header descriptions
ID_REF
VALUE
GeneSpring 7.3.1 was used to load and median-normalize raw gpr files. Matrix of values was copied directly from GeneSpring into Excel as double-precision values.