title: Allergochip for asthma diagnosis
status: Public on Jan 03 2012
contact.city: Perugia
contact.first: tania
contact.last: dottorini
contact.address: via del Giochetto
contact.department: Dept. Exp.Medicine and Bichem. Sci.
contact.country: Italy
contact.institute: University of Perugia
organism: Dermatophagoides pteronyssinus
manufacture_protocol: The array contained 103 allergens printed in duplicate onto aldehyde-activated glass microscope slides (CEL Associates) using high-speed robotics (Microgrid Compact; Biorobotics). Arrays were printed at 23°C/60°C humidity and stored overnight inside the printing cabinet. Allergens (provided from Allergopharma) were initially reconstituted in PBS pH7.4 (reconstitution buffer) with a final concentration ranging form 0.4 to 40 mg/ml and after that spotted onto the arrays in the following spotting buffer PBS pH7.4, glycine pH2.4, Borate pH9.4, glycerol 10%, DTT 5mM, SDS (0.2%; 0.05%), Tween 20 (0.01%). Allergens spotting concentration ranged from 0.008 to 3 mg/ml.
  Goat anti-human IgE  was spotted on 4 spots for positive control. Spotting buffer was printed on 46 spots with the aim of negative control. Internal calibration curve included 7 spots (duplicate).  103 spot data (duplicate) were printed on the array. Therefore, a total of 270 spot were included in the array.
  A detailed description of the sample (allergens, stock and spotting concentration, spotting buffer) is provided upon request.