However, one set of blood samples from the Zoological Museum University of Copenhagen (n = 45) were not stored in lysis buffer and were kept in −80°C freezers.
Briefly, Weck-Cel sponges (Eyetec Ophthalmic products, Altomed Ltd ,UK) were pre-wet in disposable tubes with 50 microL of sterile PBS buffer, and were kept under flow hood, at room temperature until sampling.
Samples were vortexed till thawing the extraction buffer and were kept on ice for 20 min.
After production and purification as described above, all rat and human full-length or recombinant TrxR and SecTRAP preparations were subjected to desalting with NAP-5 columns (GE) for buffer change to 50 mM Tris-Cl, 2 mM EDTA, pH 7.5 (TE-buffer) and were kept in −20°C at a concentration of approximately 1 mg/ml until use.
Clinical samples were collected in phosphate buffer solution and were kept in this solution less than 3 min, followed by immediate immersion in the tissue stabilization solution RNAlater (Ambion, Applied Biosystems, Foster City, CA, USA).
Jurkat T-cells (∼2×10 2.5×10) were harvested by centrifugation (550 g for 5 min at 4 °C), washed once with buffer A (containing 20 mM Hepes, pH 7.4, 140 mM NaCl, 5 mM KCl, 1 mM MgSO4, 1 mM CaCl2, 1 mM NaH2PO4 and 5.5 mM glucose), resuspended in 1 ml of buffer A and were kept at room temperature for 10 min.
The samples were prepared using phosphate buffer and 2%% glutaraldehyde and were kept overnight at 4 °C for fixation.
All stock buffers were stored at 4°C and were kept for 1 month.
For mapping with DMS, CMCT, and kethoxal, RNRspBs was annealed in 20 μL of a folding buffer as described above and was kept at 22 or 4 °C.
Treatment of different rabbit groups was as follows: group (1) was injected S/C with 2 mL sterile phosphate buffer saline solution (PBS) and was kept as normal control, group (2) was injected S/C with a single dose of propolis, group (3) was vaccinated with P. multocida vaccine only, and group (4) was injected S/C with the vaccine mixed with propolis as an adjuvant.
Any remaining of the droplet was aspirated, the fibers were washed three times with the buffer and they were kept in buffer prior to any further processing.
