Briefly, logarithmically growing D. discoideum cells were washed twice in 17 mM phosphate buffer (pH 6.2) and kept as frozen pellets of 2×107 cells at −80°C.
This antibody was purchased from Abcam (Cambridge, UK), kept as frozen aliquots as recommended by the manufacturer, and used in parallel to the three antibodies we generated ourselves.
RNA was isolated using Trizol Reagent, dissolved in RNA storage solution (AM7000, Ambion/Applied Biosystems, Austin, TX, USA) and kept as frozen aliquots at -80°C.
To examine whether peptides 8.1 and 8.3 can prime CTL in human lymphocytes in vitro, PBMC of healthy donors were collected by leukapheresis, and kept as frozen aliquots.
P. putida strains, kept as frozen stock in 25% glycerol at −80°C, were streaked on Luria Bertani agar plates and incubated for one day at 30°C.
N-myristoylated cell-permeable PKC and PKA inhibitor peptides with specific pseudosubstrate sequences (myr-FARKGALRQ-amide, and myr-GRTGRRNAI-amide, respectively) were purchased from Merck (Darmstadt, Germany), reconstituted in H2O at 10 mM, and kept as frozen stocks.
Liver samples (nodules and un-inflamed normal areas), all other major organs and serum (by cardiac puncture) were collected at 3-month intervals until 15 months, for histological analysis, RNA extraction, genomic DNA and serum extraction, and were kept as frozen samples or fixed in 4% formalin.
One reality is that the prices of certain essential services, like subway fares and college tuition charges, enjoyed the Zamboni treatment for many years; they were kept as artificially frozen as the hockey ice at Madison Square Garden.
Single colonies were picked up from Columbia SB agarized plates (Beckton Dickinson, Cockeysville, MD), grown in Pseudomonas broth containing Gm, 50 μg.ml-1 and kept at -80°C as frozen stocks containing 8% glycerol.
Either suspended in liquid or kept as a freeze-dried powder, vaccines could be stored for 1 year at ambient temperature (20°C) with minimal loss of immunogenicity in mice.
