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The pH and buffering capacity of stimulated saliva are measured using Saliva-check BUFFER kits (GC, United States of America) [ 51, 52].
The library was immobilized with Strepavidin beads (Library Immobilization Bead and Buffer Kits from 454 Roche) and single stranded with 0.125N Sodium Hydroxide.
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Ca2+ titration was done with premixed Ca2+ buffers (Calcium Calibration Buffer Kit #2 and #3, Invitrogen).
Bacterial extracts were processed for protein purification using a His-Bind Resin column and buffer kit (Novagen).
Cross-linked cell lysate was prepared using a RIPA buffer kit (Santa Cruz) as per the manufacturer's instructions.
Ca2+ calibration of Fura-2 fluorescence was performed in vitro using the Ca2+ calibration Buffer Kit #2 (Invitrogen-Molecular Probes, Eugene, OR).
Intracellular staining for FoxP3, CTLA-4 and cytokines was performed using the eBioscience fixation/permeabilization buffer kit following manufacturer's instructions.
The fusion protein was purified by affinity chromatography with the HisBind resin and buffer kit, in accordance with the manufacturer's instructions (Novagen/Merck Chemicals, Nottingham, UK).
For the 5' end regions with high GC contents at the wx gene, LA Taq™ with a GC Buffer kit (Takara) was used.
Calibration, enabling linking fluorescence intensity with intracellular concentration, was performed using the calibration buffer kit, exposing calcium probe-loaded cells to buffers which free Ca2+ concentration was set between 0 and 39 µM with appropriate EDTA concentrations.
Pretreated cells were re-suspended in assay buffer (Kit components) and seeded (1.5x10) in each well.
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