Incubation solutions

Eight samples from each set (A and D) were split into two groups, one incubated in 0.15 M phosphate buffered solution (PBS) and the other in PBS containing 0.1% of the phase transfer catalyst, tricapryl methyl ammonium chloride (Aldrich Aliquot 336) (PBS, catalyzed), both at pH 7.4. The samples were suspended in centrifuge tubes with 10 mL of incubation solution by nylon string and the tubes were placed in a shaker bath at a constant temperature of 37°C. Periodically, the samples were taken out of the tubes, placed on lint free paper towels, allowed to dry for exactly one hour, weighed and placed in fresh incubation media. The total time of the study was 48 weeks (330 days). 

The protocol of experiments includes the following a) the incubation in complete incubation solution ( standart incubation , 1-st channel of flow... 

The incubation conditions for trypsin and pepsin were chosen after preliminary tests showed that measurable amounts of collagen were released into the incubation solutions. In trypsin incubations, degradation is fastest on day 1 and decreasing in velocity thereafter. Slices digested extensively by pepsin were fragile. Infermittent changes of pepsin solutions were therefore avoided. 

Salicylic Acid Absorption. The apical 5 cm of the primary and two seminal roots from each of three plants were out into 1-cm segments to form an experimental unit (ca. 0.08 g). Incubation solution cc ijjtalned 0.5 mM KCl, 0.25 mM CaSOjj, 0.5 mM salicylic acid, 10 nCl/mL [ C]-sallcyllc acid, with 25 mM Tris and 25 mM Mes buffers mixed to obtain pH 6.5. Because the salicylic acid was dissolved in absolute ethanol, the final concentration of ethanol in the incubation solution was 1 (v/v). Root segments were transferred to test tubes containing 10 mL continuously aerated incubation solution. After the predetermined absorption time, segments were collected from the incubation solution by rapid filtration on Whatman No. 2 filter paper. 

All determinations are performed in duplicate. The blank is 10 pi BSA solution + 10 pi MU-NeuAc substrate pH 4.3 in a well of a 96-well plate. The sample comprises 10 pi homogenate (diluted until the protein concentration is 3 mg/ ml) + 10 pi of substrate. Incubate the samples for 1 h at 37°C on a heat block. Add 200 pi carbonate buffer and read the fluorescence. For calibration, 25 pi 4-MU standard solution (750 pmol) is mixed with 200 pi carbonate buffer and measured. For / -galaclosidase, the homogenate is diluted until the protein concentration is 0.3 mg/ ml. The incubation solution is as follows 10 pi + 20 pi MU-Gal. 

For sealing membranes into bags it is easiest to cut two sheets of plastic and place one underneath and one on top of the membrane Heat seal around three sides to make a bag, add the incubation solution, remove any air bubbles and seal the fourth side... 

A.M. Oliveira-Brett and V.C. Diculescu, Electrochemical study of querce-tin-DNA interactions. Part I—Analysis in incubated solutions, Bioelectrochemistry, 64 (2004) 133-141. 

Separate the MB with the immobilized BC-A from the incubation solution, decant and wash sequentially with 100 pL of TT, 100 pL of TTE and 100 pL of TT buffers with the appropriate magnetic separation steps and then decant and resuspend in 50 pL of hybridization solution. The suspension of MB-modified with BC-A is ready for the hybridization. 

Increasing concentrations of the antigen (fungicide) are incubated with a fixed concentration of antibody. With increasing antigen concentration, more of the antibodies in the incubation solution will be complexed and inactivated. 

FIG. 7 The measured relation between ArpH7 and concentration of lysozyme in the incubation solution for a pH step from pH = 4 to 9 and an incubation time of 30 min. 

Continuous methods do not require a separation step prior to detection. For assays using this method, the substrate and product must differ in some property such that either one may be measured directly in the incubation solution. For example, the activity of an enzyme catalyzes the conversion of 4-nitrophenyl phosphate (4NP), a colorless compound, to 4-nitrophenol, which is yellow and has an absorption maximum at 510 nm. Since the substrate does not absorb in this region of the spectrum, the reaction can be carried out... 

Reaction mixtures contained substrate (ALA) and buffer the reaction was started by the addition of lysate. TCA was added to terminate the reaction. The incubation solution was clarified by centrifugation, and a constant volume removed and mixed with the internal standard. A known volume was injected for analysis. Figure 9.59 compares the profiles of two samples after incubation with enzyme and with blank. The appearance of the reaction product PBG is observed in the test profile (Fig. 9.59A) only. 

After 7 min, aspirate the supernatant gently and add 50. 1 of preincubation or incubation solution. 

Solubility solution conditions are important. The pH-solubility profile is a function of the intrinsic solubility of the neutral form, with the solubility of the ionized species (protonated for bases and deprotonated for acids) being typically much higher than that of the neutral species. Therefore, pKa as well as concentration of DMSO present in the final incubated solution needs to be considered. 

Common for all the membrane-, cell- and tissue-based systems used to determine drug permeation, is the need for an incubation solution preserving the viability, functionality and/or integrity of the membranes, cells or tissue. It is also important that the incubation solution does not influence the permeability characteristics of the epithelial membrane or lipid bilayer and that the drug compound is chemically and physically stable and sufficiently soluble in the buffer. In all models, aqueous incubation solutions are used, often with slight modifications depending on the purpose of the study. 

AEC stock solutions (0.4% in DMF) are stable at room temperature. The incubation solution is prepared by adding 0.5 ml AEC stock solution to 9.5 ml 50 mM acetate buffer, pH 5.0, followed by 1-10 pi 30% H2O2. The solution is filtered onto the sections and left for 3-10 Ynin at room temperature. This reaction is generally weak. 

When the immobilization was complete separate magnetically the resulting MB/CFA conjugate (MB with the immobilized CF-A), from the incubation solution by placing the tube on the magnet for 1 min. 

Separate the formed MB/anti-human IgG from the incubation solution and wash three times with 150pL of B W buffer. 

Add 15% of the total culture volume (v/v) of PEG/NaCl solution to each centrifuge bottle. Mix very thoroughly by multiple inversions and incubate solutions at 4°C overnight to allow the phage to precipitate. 

Incubation in protein-free SBF (HBSS) for up to 12 weeks resulted in preferential dissolution of thermal decomposition products and ACP with a concurrent increase of the HAp content to between 85% and 90% (Table 6.6). From the incubation solutions secondary HAp precipitated. Depending on the coating type, very different precipitation kinetics were observed that resulted in morphologically different layers of secondary HAp. The precipitation kinetics could be followed by measuring the change of the relative rate of the phosphate ion concentration in solution (Figure 6.3). 

Incubations of microscope slides with small amount of incubation solution must be done in a closed box with a source of water vapor (moist paper towels) to prevent the small volume of solution from drying. While it is possible to buy boxes, most researchers like to make their own with recycled plastic boxes from lab products. Incubation times are generally 24-48 h. Evaporation of the small volumes is a major problem. Agitation, while needed to shorten the incubation time, is not used because of the danger that the incubation solution will run outside the rings. 

Fig. 4.5 Incubating tissue sections. For incubations of tissue sections on microscope slides, wells are made around sections, (a) To hold the incubations solutions in wells around sections, a pen with hydrophobic material is used to paint a rectangle around the section, (b) Section through the microscope slide showing the line made by the pen and the tissue section, (c) The incubation solutions are held in place by the hydrophobic lines...

The fourth category. Incubation Solutions, includes information on the species of the normal blocking serum and the detergent. 

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