Incubation temperature

Manufacturer and system Throughput Samples per hour Tests Number of resident methods Amount of sample needed per test, p.L Incubator temperature, °C Optical a system Distinctive features... 

The medium in each vessel is then inoculated with a heavy aqueous suspension of spores of a strain of Actinomyces griseus, and the inoculated media are maintained at an incubation temperature of 22° to 28°C for 10 days. The growth is then filtered off and the filtrates are combined for further treatment. 

Cultivation conditions Spore of the fungus was inocvilated into 20 g solid substrates composed of wheat bran, rice bran and rice husk in a 9x14 inch plastic bags (Figure 2). Different ratios of wheat bran, rice bran and rice husk were carried out as well as different initial pH, moisture content and incubation temperature were concerned. The fungus in the solid substrates was incubated for 6 days. 

Figure 5 Pectinase activity in solid substrates of wheat bran, rice bran and rice husk at different ratios of substrates. Initial moisture content 66 % Incubation temperature 32°C Initial pH 5.7...

Figure 6,7 and 8 showed the results of the pectinase activity when produced in the solid substrates containing wheat bran, rice bran and rice husk in the ratio of 6 12 2. The highest activity obtained when the strain was grown on the solid substrates with 58 % initial moisture content, pH adjusted to 5.7 and incubation temperature was at 32°C. Under these conditions, the highest activity of the enzyme that could be obtained from Rhizopus sp. 26R was ca. 700 units of enzyme activity per gram of solid substrates. 

Wu Q, DL Bedard, J Wiegel (1997a) Effect of incubation temperature on the route of microbial reductive dechlorination of 2,3,4,6-tetrachlorobiphenyl in polychlorinated biphenyl (PCB)-contaminated and PCB-free freshwater sediments. Appl Environ Microbiol 63 2836-2843. 

Andersen et al. (1996) and Andersen (1995) have studied the effect of temperature on the recombinant protein production using a baulovinis/insect cell expression system. In Tables 17.15, 17.16, 17.17, 17.18 and 17.19 we reproduce the growth data obtained in spinner flasks (batch cultures) using Bombyx mori (Bm5) cells adapted to serum-free media (Ex-Cell 400). The working volume was 125 ml and samples were taken twice daily. The cultures were carried out at six different incubation temperatures (22, 26,28, 30 and 32 TT). 

GULF TOADFISH, Opsanus tau Isolated hepatocytes acclimatized to 18 or 28°C, then exposed to BaP at 18, 23, or 28°C for about 2 h Isolated gill cells acclimatized to either 18 or 28°C for 3 weeks and exposed for 8 h to 1-100 mg BaP/L at incubation temperatures of 18, 23, and 28°C Dose-dependent increase in metabolite production. 23 Rapid metabolism of BaP independent of temperature Accumulation followed a dose-concentration gradient 24 uptake rate was higher for cells acclimatized to 18°C than those acclimatized to 28°C at all incubation temperatures. When cells were exposed to BaP at the respective acclimatization temperatures, uptake rates were similar... 

Raising the incubation temperature from 45 to 57°C did not bring about a pronounced increase of the hydrogen-driven pMMO activity. This preliminary observation indicated in vivo heat stability of the hydrogenase and pMMO activities. The temperature dependent difference in the solubility of hydrogen may also explain the small activity difference, particularly as similar results were obtained for the hydrogen-driven sMMO activity. 

A few other official antibiotics in BP (1993) may also be assayed by adopting the method stated above, but using specific micro-organism, definite final pH of the medium, pH of the phosphate buffer, potency of solution (U per ml) and the incubation temperature. A few typical examples are given in Table 20.1 below ... 

S. No. Antibiotic Micro-organism Medium Final pH Phosphate Buffer pH Potency of Solution U per ml Incubation Temperature (°C)... 

Several modifications of incubation conditions have neither stabilized the system nor enhanced activity. Acetone and methanol have been used as substrate carriers without affecting activity. Similarly, addition of NADH to the incubation media did not effect epoxidation. The enzymatic nature of the system has been confirmed by use of heat treated homogenates (100 C, 1 min). Incubation temperatures of 8, 20, and 30 resulted in progressively greater epoxidation rates and provided no evidence of heat lability. Thus, at this time it is not possible to identify a superior enzyme source for comparative studies in spite of the fact that in vivo measurements indicate oxidative metabolic activity in living mussels. 

One AHH fluorescence unit equals the fluorescence intensity of a 3 ug/ml quinine sulfate 2H20 in 0.1 N sulfuric acid solution (excitation A 425 nm, emission a 555 nm). 7-Ethoxyresorufin deethylase activity was assayed essentially as described by Burke and Mayer (13) at a 7-ethoxyresorufin concentration of 2 yM, a final pH of 7.8, and an incubation temperature of 30° (14). 

The effect of pH on in vitro aldrin epoxidase activity was established over a pH range 6.5-8.5 (Figure 1). A pH of 7.5 was used as optimum. The effect of temperature on in vitro aldrin epoxidase activity was determined over a range of 20°-40°C (Figure 2). An optimum incubation temperature of 30°C was used. The maximum epoxidase activity was attained at a Tris-HCl buffer concentration of 5.0 X 10"1 M (Figure 3). 

Since active transport mechanisms require energy, the incubation temperature during the assay plays a crucial role. At 4°C, the fluidity of the cell membrane is reduced, the metabolism of the cell is downregulated, and energy-dependent transport processes are suppressed. Consequently, the amount of cell-associated target system refers mainly to the cytoadhesive fraction. In contrast, incubation at 37°C increases the fluidity of the cell membrane and the metabolic activity to an optimum, so both cytoadhesion and cytoinvasion occur at the same time. Thus, the uptake rate can be calculated from the difference in signal intensity measured upon incubation at both respective temperatures. 

During chase incubation at 4°C, no significant change of the cell-associated fluorescence intensity should be detected over time. Because of the reduced incubation temperature, the metabolism of the cells is minimized, resulting in an inhibition of active uptake processes. Upon subsequent addition of monensin, the fluorescence emission signals should not be altered as well. In doing so, any direct influence of monensin on the quantum yield of the fluorescein label can be excluded [25],... 

The BCA assay has a working range of 20 to 2000 Lig/ml. If the target protein is in a dilute aqueous formulation, the concentration can be determined with the micro BCA assay. The BCA protocol is modified by increasing the BCA concentration, incubation time, and incubation temperature. These modifications permit the detection of BSA at 0.5 Lig/ml. The major disadvantage of these modifications is that the presence of interfering substances decreases the signal-to-noise ratio and thus the sensitivity of the assay. 

The cells are permitted to "plant" to the ECM and adjust to the incubator temperature (37°C) and C02 concentration. Then test compounds or controls (both in 0.1% dimethyl sulfoxide, DMSO) are added to the test wells. The cells are then incubated overnight, and the indicator dye Alamar blue10 is added. This noncytotoxic dye reacts to mitochondrial redox reactions and is measured fluorometrically. Cell metabolic activity is determined starting at 3 h after the dye is added and daily thereafter. 

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