Chromatography was performed on a guard column (SecurityGuard® Guard Cartridge Kit; Phenomenex, Torrance, CA, USA) and on a C18 column (Brownlee C18®, 5 μm, 150 × 4.6; PerkinElmer) at 37°C.
The resulting solution was injected (5 μl) onto a 1200 series LC system (Agilent Technologies), consisting of 1200 series binary pump, autosampler, vacuum degasser, column compartment and control module, with a Luna 3 μm C18(2) 100 Å, 100 mm × 2.00 mm column and a SecurityGuard cartridge kit (Phenomenex, Macclesfield, UK) as a guard column.
Samples were subsequently cleaned with a cation exchange Cartridge Kit (Applied Biosystems).
Replicas of mouse dorsal skin were obtained using a Repliflo Cartridge Kit (CuDerm Corp., Dallas, TX, USA).
After extraction, the ICAT-labeled peptides were purified using the ICAT Avidin Buffer Pack and Avidin Affinity Cartridge kit (Foster City, CA) according to the manufacturer's guidelines.
The amount of irradiation was progressively increased from 60 mJ/cm per exposure at week 1 (1 minimal erythematous dose =60 mJ/cm) to 90 mJ/cm at week 7. Replicas of mouse dorsal skin were obtained using the Repliflo Cartridge Kit (CuDerm Corp., U.S.A).
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To reduce non-specific reactions and gain a higher sensitivity, high molecular weight proteins contained in the sera were removed by Multiple Affinity Removal Spin Cartridge Reagent Kit (Agilent Technologies, Santa Clara, CA, USA) and used for the serum samples.
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68Ga was retained on a cation exchange cartridge (Reagent and Hardware Kit SC-01, ABX, Radeberg, Germany) in a GRP Module (Scintomics, Lindach, Germany).
This mixture was pipetted into a tube containing a filter cartridge supplied with the kit.
