Phosphate-buffered saline PBS

Adsorption of t-PA to process equipment surfaces consisting of either stainless steel or glass was minimized by adding the detergent polyoxyethylene sorbitan monooleate (Tween 80) to the semm-free culture conditioned media at 0.01% (vol/vol). The equipment was also rinsed, before use, with phosphate buffered saline (PBS) containing 0.01% Tween 80. Hydrophilic, plastic equipment was used whenever possible. AH buffers were sterile filtered. Sterile filtration of Hquids and gases is usually carried out using 0.2 or 0.45 p.m filters. 

FIGURE 9.4 Circular dichroistn (CD) spectrum of dilute solution (0.24 mg/mL) of recombinant resilin in phosphate-buffered saline (PBS). (From Whelan, A.J. and Robinson, A.J., unpublished data). 

FIGURE 9.6 DSC of (a) recombinant resilin in water showing no enthalpic events, (b) bovine serum albumin in phosphate-buffered saline (PBS) showing denaturing occurring at 62°C, and (c) wool fiber in water showing denaturing of the a-helix at 145°C (Endotherm up). 

FIGURE 9.8 Typical stress-strain plots for a strip of recombinant resilin tested in phosphate-buffered saline (PBS). Sample cycled to 225%, showing resilience of 97% (solid curve) and later tested to failure showing extension at break of 313% (dotted curve). (FromElvin, C.M., Carr, A.G., Huson, M.G., Maxwell, J.M., Pearson, R.D., Vuocolo 1, T., Liyon, N.E., Wong, D.C.C., Merritt, D.J., and Dixon, N.E., Nature, 437, 999, 2005.)... 

FIGURE 9.9 Small dumbbell-shaped sample of recombinant resilin prepared with embedded fine stainless steel mesh tabs (left) and being tested in phosphate-buffered saline (PBS) (right). 

In a subsequent study, the effect of reducing the ELP molecular weight on the expression and purification of a fusion protein was investigated. Two ELPs, ELP [V-20] and ELP[VsA2G3-90], both with a transition temperature at 40°C in phosphate-buffered saline (PBS) containing 1 M NaCl, were applied for the purification of thioredoxin. Similar yields were observed for both fusion proteins, resulting in a higher thioredoxin yield for the ELP[V-20] fusion, since the ELP fraction was smaller. However, a more complex phase transition behavior was observed for this ELP and therefore a selection of an appropriate combination of salt concentration and solution temperature was required [39]. 

Figure 8. Standard curves for PbTx-2 ( ) and PbTx-3 ( ) in the brevetoxin radioimmunoassay. Lower detection limits are 0.2 — 0.5 ng in phosphate-buffered saline (PBS). Standard RIA conditions [ H]PbTx-3 = 1 nM antiserum dilution = 1 2000 sample vol. = 1 ml buffer = 0.1 M PBS, pH = 7.4.

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... 

Instruments, Inc., Rockville, MD) was added, and the proteins were fractionated using a 10-channel RIEF apparatus. The proteins in each of the 10 fractions were then dialyzed against large volumes of phosphate-buffered saline (PBS, pH 7.4). Immunoelectrophoresis and a radioimmunoassay (RIA) for PCP were used to determine the RIEF fractions with the greatest purity and anti-PCP activity. 

The disassembly rate of dendritic molecules 18 and 19 was evaluated in phosphate buffered saline (PBS, pH 7.4) in the presence and absence of PGA. The release of tryptophan was monitored by a reverse-phase HPLC at a wavelength of 320 nm. The results are presented in Figs. 5.11 and 5.12. No disassembly of either system was observed in the buffer without PGA (data not shown). In the presence of PGA, dendritic molecule 18 disassembled to release tryptophan within approximately 4 days (Fig. 5.11), whereas dendritic molecule 19 released its tryptophan tail units within 40 min (Fig. 5.12). 

Although several types of fluorescent beads were proposed as a microscopic fluorescence standard 30 years ago,2 beads have not been used as a proteinembedding matrix for routine IHC on FFPE tissue. We recently tested primary coated beads ( Dynabeads, Dynal, New York) that are coated with a goat anti-mouse antibody on the surface of the beads. In the first experiment, a monoclonal antibody to cytokeratin 7 (DAKO, 50pL/34.5pg) was bound to the beads by incubating with the beads (150 pL at a concentration of 109 beads/1 pL) at 4°C in a cold room with an automatic shaker for overnight. Incubation was followed by three phosphate-buffered saline (PBS) washes,... 

Figure 4. AFM micrograph of a saturated monolayer of antibodies against p2-microglobulin measured in phosphate buffered saline (PBS) solution using tapping mode. The dark window shows the underlying polystyrene surface obtained by wipping off the antibodies.

Although the signal reagent can be used over a wide pH range, the optimal pH was found to be 5-7. Most of the results described in this chapter were obtained using a pH of 5.4 [phosphate-buffered saline (PBS), 10 mM],... 

The protein A (pA), antihuman serum albumin (a-HSA, M 150 kD), and human serum albumin (HSA, M 65 kD) were provided by Paradocs BV (Tiel, The Netherlands). The Herpes Simplex Virus type 1 (HSV-1) and anti-HSV-1 gG glycoprotein G monoclonal antibody (a-HSV-1 gG) were purchased from Virusys Corporation (Marriottsville, MD, USA). Bovine serum albumin (BSA, M 50 kD) was purchased from Sigma-Aldrich Chemie BV (Zwijndrecht, The Netherlands). Synthetic surface protein of Hepatitis-B virus generated in Hep-G2 cell-line (HEP G2, M 25 kD) was provided by BioMerieux BV (Boxtel, The Netherlands). Phosphate buffered saline (PBS) was used for all experiments. 

Treat sections on the grids with phosphate-buffered saline (PBS) at pH 7.4 containing 0.05% Triton X-100 for 15 min at room temperature. 

Each cell monolayer is rinsed with a minimum of 20 ml phosphate buffered saline (PBS) and then 20 ml of treatment medium is carefully added. The flasks are incubated for 3 h at 37°C in a C02 incubator. 

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. 

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