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Separated spots were located using an ultraviolet analyzer with 254 nm wavelength (WFH-203B, Shanghai Jing Branch Industrial Co., Ltd., China).
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The injection spot was located using a 10× objective.
For each calibration image of the projected spots, the centroid of each spot was located using the algorithm described in Section 2.2.
The microarray spots were located by using the GenePix Pro 6.0 software package (Axon Instruments, Foster City, CA) together with the array list file [ 67].
Spots were located and quantified with the QuantArray 3.0 software (Packard BioScience) using the histogram quantification method to determine the mean intensity of each spot.
Protein spots were located and excised with an EXQuest™ spot cutter (Bio-Rad).
The hot spots were located in stable hairpin structures [ 4].
These HK-PPD spots were located between 10 - 15 kDa and possessed approximate pIs of 6 - 8. While these spots could represent additional cytoplasmic proteins released into HK-PPD through the autoclave process, we did not detect antibody recognition to these protein spots in corresponding locations using Western blot analysis (data not shown).
Major protein spots were located in acidic range (pH 4 7) migrating at 30 60 kDa.
Radioactive spots were located by autoradiography, scraped off and counted by liquid scintillation.
Significant breast cancer hot spots were located near known groundwater plumes and the Massachusetts Military Reservation.
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