[Optimization of the conditions for protein chip preparation].
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.
Jiang
Ling-xiao
LX
Department of Epidemiology, Southern Medical University, Guangzhou 510515, China.
Guo
Zhao-biao
ZB
Chen
Ze-liang
ZL
Wang
Jin
J
Wazng
Hong-xia
HX
Yu
Shou-yi
SY
Yang
Rui-fu
RF
D040081:Protein Array Analysis / Q000592:standards*; D011786:Quality Control; D012680:Sensitivity and Specificity