Anti-GST polyclonal antibody

The cyanobacterial circadian clock is the most well-understood and simplest biological time-keeping system. Its oscillator consists of three proteins: KaiA, KaiB, and KaiC. When combined together in a test tube, the Kai proteins produce a free-running 24-h cycle of rhythmic auto-phosphorylation and auto-dephosphorylation. To generate a robust circadian rhythm of the in vitro reaction mixture, KaiC, the core oscillator protein, must be purified with an untraditional approach, since even the smallest amount of impurity can hinder its post-translational activities. Until recently, series of fast protein liquid chromatography (FPLC) columns (glutathione S-transferase (GST), anion exchange (Q), and desalting columns) have been used to purify the oscillator proteins, often requiring laborious elution processes. Although the common methodology has already been established, whether the purified KaiC can produce robust oscillations remains to be verified. Here we emphasize the significance of eliminating the Q step and lengthening the step of removing the non-specifically bound impurities on the GST column for generating a rhythmic KaiC phosphorylation in vitro. These findings demonstrate the potential for shortening the amount of time and effort it takes to purify proteins without compromising its quality.

The dusky grouperEpinephelus marginatusis predator fish subjected to be impacted due to the contamination of their habitats. A viable source of metal contamination, i.e., copper (Cu), in this species is the ingestion of contaminated food. The objective of this work was to verify the toxic effects of Cu contaminated feed in dusky grouper. A 15 days trial was conducted with three treatments: control, 1 g and 2 g Cu/kg of fish feed. After the trial, the gut was analyzed for Cu concentration and the liver for SOD and GST activity. The Cu concentration in the intestinal tract was significantly greater in fish from contaminated treatments when compared with control. The SOD was significantly lower in contaminated fish, and the GST did not show differences among treatments. Copper showed to be toxic for the species, as evidenced by gut accumulation and suggested by SOD response.