Hydroxystilbamidine bis(methanesulfonate)

Kinase experiment:

Silkwork larvae are used in this study. Larvae on the fourth or fifth day of the fifth instar (~4.2 g body weight) are injected with 35 μL of a solution of 10 mg/mL cyclobeximidein H2O. After 5 min, the animals are immobilized in ice, and posterior silk glands are dissected and washed in ice-cold 0.15 M NaCl, 0.015 M Na citrate, 100 μg/mL cycloheximide. Washed glands from two larvae are placed in a homogenizer containing 4.7 mL of 40 mM triethanolamine-HCl, pH 7.5, 0.15 M sucrose, 0.1 M KCI, 3 mM MgCl2, 2 mM reduced glutathione, 10 μg/mL cycloheximide, 750 μg/mL Escherichia coli tRNA, and an appropriate concentration of RNase inhibitor (sodium heparin, 1.5 μg/mL orHydroxystilbamidine bis(methanesulfonate), 1.5 mM)[1].

Animal experiment:

Mice (six per group) are given various doses of Hydroxystilbamidine bis(methanesulfonate) (HSB) 3, 2, and 1 day before antigen. Other groups are given Hydroxystilbamidine bis(methanesulfonate) 1 or 2 days after the injection of antigen. Another group of mice receive antigen and Hydroxystilbamidine bis(methanesulfonate) simultaneously. A control group receives only antigen. The antigen dose consists of 2×108 sheep erythrocytes (SRBC). Four days after the injection of SRBC, the mice are sacrificed and spleens are removed and assayed for plaque-forming cell (PFC) by the plaque assay[2].

References:

[1]. Lizardi PM. Isolation of giant silk fibroin polysomes and fibroin mRNP particles using a novel ribonuclease inhibitor, hydroxystilbamidine. J Cell Biol. 1980 Oct;87(1):292-6.
[2]. Folds JD, et al. Immunosuppression by hydroxystilbamidine isethionate, a lysosome-stabilizing, anti-proteolytic, antifungal drug. Infect Immun. 1975 Mar;11(3):441-4.