Phosphate-buffered saline, solution preparation

Measurement. The measurement apparatus for the in vitro test of this sensor is shown in Figure 4. The sensor was dipped into the flask containing 100 ml of phosphate buffered saline solution, pH 7.4, and the output was measured with respect to stepwise changed glucose concentrations of 0 to 2000 mg/dl under the oxygen tensions of 5 to 21%, which prepared by mixing air and N2 gas. 

Carrier solution is phosphate buffered saline (PBS), prepare by adding 7.6 g ofNaCl per 1L of the phosphate buffer. Optionally, it might be useful to add detergents (e.g., Tween-20, P1379, 0.01%) and inert protein (e.g., 250 Xg/mL bovine serum albumin, 05477) to suppress nonspecific binding of proteins and other molecules from samples. 

Figure 4 Estradiol flux through human stratum comeum when the drug is applied oedu-sively in liquid or gel state vesicles prepared from polyoxyethylene alkyl ethers and cholesterol. Ail formulations are saturated with respect to estradiol. (A) Plus estradiol applied in PBS , estradiol applied in liquid state vesicles prepared from decaoxyethylene oleyl ether (Q.gEOjo) and cholesterol , estradiol applied in phosphate buffered saline solution (PBS) after pretreatmem of vesicles prepared from C,.,EOjo and cholesterol. (B) Plus estradiol applied in PBS , estradiol applied in gel state vesicles prepared from trioxy-ethylene alky octadecyl ether (CitEOr) and cholesterol . estradiol applied in PBS after pretreatmem of vesicles prepared from C,i,EOj and cholesterol.

Washing solution phosphate-buffered saline (PBS). Prepare 10 X stock with 1.37 M NaCl, 27 mM KCl, 100 mM Na2HP04, 18 mM KH2P04 (adjust to pH 7.4 with HCl if necessary) and autoclave before storage at room temperature. Prepare working solution by dilution of one part with nine parts water. 

The specifications and standardization include raw materials, preparation method of the standard solution, concentration of proteins, and the main band on SDS-PAGE. The outline of the procedure for preparation of the calibrators is shovm in Eig. 4.2. Table 4.5 shows the raw materials and the preparation method of the initial extract. To prepare the calibrators, the raw materials are extracted by the standard solution containing SDS and mercaptoethanol. The initial extract is prepared by centrifugation and filtration of the extract. The diluted extract is then prepared by 10-fold dilution of the initial extract with phosphate-buffered saline (PBS pH 7.4). The protein concentration of the diluted extract is assayed using the 2-D Quant kit (Amersham Bio Sciences). The standard solution is then... 

Preparations of PEG-modified proteins. A. SC-PEG (1 g, 0.2 mmol) was added to a stirred solution of Bovine Serum Albumin (BSA) (100 mg, 1.5 x 10 6 mol) in 0.1 M sodium phosphate, pH 7.8 (60 mL). Sodium hydroxide (0.5 N) was used to maintain pH 7.8 for 30 min. The excess of free PEG was removed by diafiltration using 50 mM phosphate buffered saline. Approximately 30 amino groups of the native protein were modified as determined by trinitrobenzenesulfonate (TNBS) assay (28). The same degree of modification was obtained when the experiment was repeated under identical conditions using SS-PEG instead of SC-PEG. 

A confluent monolayer of Madin-Darby canine kidney (MDCK) cells was grown in 96-well plates. Serial tenfold dilutions in minimal essential medium were prepared from the aliquots of allantoic fluid taken from the irradiated specimen. These dilutions were applied to MDCK cells and incubated for 48 h at 36 °C in 5% C02. The cells were then washed two times for 5 min with phosphate buffered saline (PBS) and incubated for 1 h with 100 pi of 0.5 mg/ml solution of 3-(4,5-dimethyl-thiazolyl-2) 2,5-diphenyltetrazolium bromide (MTT, ICN Biochemicals Inc., Aurora, Ohio). After lh, the colored deposit was dissolved in 100 pi DMSO, and optical density in the wells was measured on plate reader Victor 1420 (Perkin Elmer, Finland). Based on the data obtained, the infectious titer of the vims was determined as a decimal logarithm of reciprocal to the dilution of the specimen causing destruction of 50% of cells. The inhibiting action of irradiation was evaluated by decreasing the vims titer. 

A Working Solution of the stain should be prepared just before use, by combining 10 ml of 0.1% Triton X-100 in phosphate-buffered saline, 2 mg RNase (DNase-free, Sigma), and 200 pi PI solution (Sigma, 1 mg/ml in distilled water). Stock solutions of Triton X-100 and PI should be stored at 4°C. Centrifuge the ethanol-fixed cells (200 x g for 5 min) and discard the supernatant, removing it completely. 

Prepare competitor protein buffer detergent solution 4 mg/mL normal goat globulin or other competitor protein, (such as fetal calf serum, bovine serum albumin, bovine plasma, and so forth), and 0.1% saponin (Sigma, St. Louis, MO) in phosphate-buffered saline (NGG-sap-PBS). 

The effects of conditioning layers of two important blood serum proteins, albumin and fibrinogen were investigated. Protein adsorption was studied using bovine serum albumin (BSA) and fibrinogen (F) from Sigma. The samples were incubated for 3 h at 37°C in solutions of albumin (1 mg/mL) and fibrinogen (0.2 mg/mL) prepared in phosphate buffered saline (PBS, 0.01 M phosphate buffer, 0.0027 M KC1, 0.137 MNaCl, pH 7.4). After the incubation period, the samples were rinsed 3 times with PBS and analyzed by the various surface characterization techniques. 

If nasal inoculation is to be performed then the animals must first be anaesthetized by intraperitoneal injection of chloral hydrate. For this, a 5.7% (w/v) solution is prepared in phosphate-buffered saline (PBS) pH 7.4, and 250 pL injected per 50 g of body weight. 

Immunoglobulin preparation, 5 mg/ml in saline Carbonate-bicarbonate buffer, 0.5 M, pH 9.5 Lysine solution, 1.0 M pH 7 Phosphate-buffered saline, pH 7.5 (PBS) Saturated ammonium sulfate Glycerol Procedure... 

Prepare an MTT stock solution of 5 mg mH (Sigma, St Louis) in phosphate-buffered saline (PBS), pH 7.5, and filter through a 0.22- x filter to sterilize and remove the small amount of insoluble residue. 

GSH stock solution 32.26 mg (Sigma) dissolved in 25 mL of phosphate buffered saline (PBS Dulbecco s ICN Biomedicals). Prepare fresh on day of assay. 

Figure 4. Preparation of the methoprene immunogen (Structure 15) by two methods a. The NHS-ester of methoprene (Structure 14) was conjugated to human serum albumin (H2N-HSA) in organic/aqueous solution, b. A water soluble active ester of methoprene (Structure 16) was prepared by the DCC coupling of methoprene-spacer acid (Structure 13) with l-hydroxy-2-nitro-4-benzene sulfonate. Reaction of compound 16 with H2N-HSA was carried out in aqueous phosphate buffered saline (PBS).

Phosphate buffered saline (PBS), pH 7.4, 10 mM sodium phosphate, 3 mM potassium phosphate, 140 mM NaCl, and 0.2 g sodium azide (to a final concentration of 0.02% w/v for prevention of bacterial growth). Before adjusting the volume (to 1 L), the pH of the solution is adjusted to 7.40. Sodium azide is not added if the buffer will be used for the preparation of liposomes for in vivo studies. 

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