title: [Optimization of the conditions for protein chip preparation].
abstract: OBJECTIVE
  To define the optimal conditions for preparing protein chips on aldehyde-coated slides.
  
  
  METHODS
  The proteins were diluted in PBS containing 40% glycerol and spotted on aldehyde-coated slides. After the spots were dried for 1 hour at room temperature, the slides were stored at 4 degrees Celsius;. Following block and rinse, the slides were used for immunoassay and the results detected with a scanner. Several key factors that might influence the results were tested, including the number of spots, concentration of protein in the spotting solution, time of immobilization and the blocking reagent.
  
  
  RESULTS
  Pre-spotting of 10 to 15 spots achieved good homogeneity of the subsequent spots on aldehyde-coated slides. The protein immobilized at 4 degrees Celsius; for 24 to 48 h showed higher fluorescence intensity and clearer images, and the slides blocked with 3% BSA produced the lowest background signal. The concentration of protein in the spotting solution significantly affected the fluorescence intensity.
  
  
  CONCLUSIONS
  To ensure good results in preparing protein chips on aldehyde slides, pre-spotting of 10-15 spots can be necessary followed by immobilization at 4 degrees Celsius; for 24-48 h and 3% BSA blocking.
authors.affiliation: Department of Epidemiology, Southern Medical University, Guangzhou 510515, China.
authors.forename: Ling-xiao
authors.initials: LX
authors.lastname: Jiang
authors.forename: Zhao-biao
authors.initials: ZB
authors.lastname: Guo
authors.forename: Ze-liang
authors.initials: ZL
authors.lastname: Chen
authors.forename: Jin
authors.initials: J
authors.lastname: Wang
authors.forename: Hong-xia
authors.initials: HX
authors.lastname: Wazng
authors.forename: Shou-yi
authors.initials: SY
authors.lastname: Yu
authors.forename: Rui-fu
authors.initials: RF
authors.lastname: Yang
mesh_terms: D040081:Protein Array Analysis / Q000592:standards*; D011786:Quality Control; D012680:Sensitivity and Specificity