Phosphate-buffered saline/ tween

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. 

Enzyme-linked immunosorbent assay (ELISA). Li and Li developed an ELISA procedure for imidacloprid to determine its residues in coffee cherry and bean extracts. A 25-g amount of sample extracted with 300 mL of methanol and 1% sulfuric acid (3 1, v/v) for 3 min. An aliquot of the sample extract (0.5 mL) is mixed with 1 mL of water and a gentle stream of nitrogen is used to evaporate methanol. The solution is then extracted with 1 mL of ethyl acetate, the extract is reconstituted in 1 mL of PBST (phosphate-buffered saline containing 0.05% Tween 20) and competitive ELISA is performed to quantify imidacloprid in the extract. Eor methanol extracts of coffee cherries and beans fortified with imidacloprid at 0.5 mgL recoveries of imidacloprid by the ELISA method were 108 and 94, respectively. 

Toxicity Tests. Assays for toxicity were conducted on ICR female mice weighing approximately 20 g each. Animals were maintained on Wayne Laboratory Animal diets (Lab-Blox) and water, ad libitum. A known quantity of toxic extract was dried under a stream of dry nitrogen and placed in a vacuum dessicator overnight. The dried extracts were dissolved in phosphate buffered saline containing 5% Tween 80 and administered intraperitoneally. Control animals received an equal volume of the vehicle. Lethality was assessed at kQ h. 

Rinsing solution IX phosphate-buffered saline (PBS) (pH 7.4) with 0.05% (v/v) Tween-20. 

Fig. 3. Schematic of staining process of SARS-CoV immunochip. (1) Spotting A high-precision robot transfers the samples, SARS-CoV proteins, and glycans of various complexities, from 96-well plate to nitrocellulose-coated glass slides. (2) Staining Before staining, the slides are rinsed with IX phosphate-buffered saline (PBS), and blocked with 1% bovine serum albumin (BSA)-PBS containing 0.05% NaN3 and 0.05% Tween-20. They are subsequently incubated with horse anti-SARS sera. The primary antibodies captured by microarrays are detected using biotinated anti-horse immunoglobulin (Ig)G, and visualized by Cy3-streptavidin.

PBST Phosphate-buffered saline with Tween-20... 

A monoclonal anti-digoxin antibody (mouse) used for the ELISA was produced and prepared by Sawada et al. [121]. An ELISA plate (96 well) was coated with ca. 100 ng/well digoxin-conjugated ovalbumin in 50 mM sodium bicarbonate buffer (pH 9.6) at 4 °C overnight. After washing with 10 mM phosphate-buffered saline (pH 7.2) (PBS) containing 0.5 ml/1 Tween 20 (T-PBS), the wells were blocked by PBS supplemented with 1 g/1 casein (C-PBS). Fifty pi samples serially... 

Wash the plate six times with phosphate-buffered saline (PBS), 0.05% Tween-20, and 2 mM of EGTA. 

Phosphate-buffered saline (PBS) containing 1% casein, or 1% Tween 20. 

The plate counts at To were estimated by diluting the neat broth culture 1 in 10 with Neutralised Peptone Water (NPW) containing per L 1.0 g Bacteriological Peptone (Oxoid, L37), 8.8 g Sodium Chloride (May Baker), 3.0 g Amisol 910 (Degussa) and 30.0 g Tween 80 (BDH, 560234H) and then decimally with Phosphate Buffered Saline (Oxoid). For biocide treated samples 1 mL was diluted in 9 mL of NPW, mixed and allowed to stand for 5 min (for neutralisation of the biocide). The solution in NPW was diluted decimally (0.1 mL in 0.9 mL) in PBS. Appropriate dilutions (0.1 mL) were plated out on Tryptone Soya Agar Plates (bioMerieux) and incubated at 37 °C for 24 h. 

A 10 mmol L stock coelenterazine solution was prepared by dissolving coelenterazine (Nanolight Technology, Prolume Ltd. Pinetop, AZ, USA) in methanol for use at a final concentration of 10 /tmol L". All coelenterazine solutions were stored at -20 °C and working solutions were kept on ice in the dark during preparation. Diluent buffers comprised distilled water (dH20), Phosphate buffered saline (PBS), Buffer A (10 mmol L Tris [pH 7.8], 1 mmol L EDTA, 0.6 mol L NaCl), 7H9 medium supplemented with Tween-80 with or without 10% OADC (oleic acid, albumin, dextrose, catalase), Luria-Bertani (LB) broth with or... 

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. 

Selection buffer PBS (phosphate buffered saline) diluted to lx from a lOx stock (Dulbecco s lOx PBS) and supplemented with 5mM of MgCl2, lOmM of KC1, and 0.01% (v/v) of Tween-20. 

PBSTB Phosphate buffered saline pH 7.4/0.05% Tween-20/ 1 mg/mL BSA. Dissolve 1 g bovine serum albumin ( A3912, Sigma Chemical Co, St Louis, MO) and 500 pL Tween 20 (Sigma Chemical Co.) in 1L PBS to make the correct final concentrations. 

Assay Diluents As the noncovalent interactions between antibody and analyte are influenced by pH, ionic strength, and temperature, typical assay buffers are isotonic solutions at or near neutral pH. Assay buffer and diluent formulations should not only promote analyte antibody interactions but also minimize the nonspecific interactions between the critical reagents and the variety of biomolecules in the sample matrix. Components such as BSA, HSA, and nonionic detergents are often included in assay buffers. Phosphate buffered saline (PBS) or 10 mM Tris-HCl solutions (both near pH 7.4) containing 1% BSA and 0.05% Tween-20 are common buffers that can also be used for dilution of test samples and detection reagents, as well as wash buffers. When wash buffers are used in large quantities and stored at room temperature, preservatives such as sodium azide or thimerasol are often added to increase their shelf life. It should be noted, however, that some components of the wash buffer may have adverse effects... 

Fisher Scientific. The 0.15 M phosphate buffered saline (PBS, pH 7.4) and 0.05 M carbonate (pH 9.6) buffers were prepared by using the appropriate sodium and ammonium salts, respectively. Tween 20 solutions (Fisher) were prepared as 0.5 ml of Tween 20 in 1 liter of 0.15 M PBS. 

At the sampling times, a sterile stainless steel cylinder (inside area = 3.56 cm ) was held firmly to the skin area to be sampled. Five milliliters of sterile stripping fluid, consisting of 0.1% Triton X-100 in 0.1 mol/L phosphate-buffered saline solution (pH 7.8), was added to the cylinder, and the skin area inside the cylinder was massaged in a circumferential manner with a sterile rubber policeman for 2 minutes. One-milliliter aliquots of this solution (10-fold dilution) were removed and plated—with 10°, 10, 10 10 and lO dilutions, as appropriate— in triplicate on trypticase soy agar containing 1.0% Tween 80 and 0.3% lecithin as neutralizers. 

CYP3A4 protein expression can be measured directly by immunodetection (LeCluyse et ah, 2000 Luo et ah, 2002). Microsomal protein (3 jig) is resolved by SDS-polyacrylamide gel electrophoresis (12% acrylamide). Resolved proteins are transferred to nitrocellulose membranes, whieh are ineubated in 3% bovine serum albumin in phosphate-buffered saline supplemented with Tween 20 (0.1 M, pH 7.4, 0.1% Tween 20) for 45 min to block nonspecific protein binding. Membranes are then treated with primary antiCYP3A4 antibody (Gentest, Woburn, MA), followed by horseradish peroxidase-conjugated antimouse seeondary antibody. The antibody-reactive CYP3A4 protein bands are visualized using enhanced chemiluminescence detection and quantitated by photodensitometry (Desai et al., 2002). 

Washing solution is phosphate-buffered saline (PBS) without addition of Tween-20. Washing requires the addition... 

Table 8 Inhibition of FMAP in f-BSA Microspheres after 60-Day Release in Phosphate Buffered Saline Containing 0.02% Tween 80 (PBST) at 37°C...

FCS)(PBS phosphate buffered saline FCS fetal calf serum) was added, incubated overnight at 4 C, and the first wash consisted of PBS/Tween. Binding of the DES to the well was maximized to 46% with the Tween wash. Anti-DES antibody (rabbit IgG) was added at a concentration of 1 900, incubated at 37 C for one hour, washed with PBS, and followed with alkaline phosphatase conjugated anti-rabbit IgG (Sigma) at a concentration of 1 3000. This was incubated for one hour at 37 C, washed with PBS, and followed with p-nitrophenyl phosphate substrate (FNP)(1 mg/ml) in diethanolamine buffer (pH 9.6). After 45 minutes in darkness, an average positive... 

Polyethyleneimine, 1% solution in water PBS (phosphate-buffered saline), lOx stock PBST (PBS + 0.1% Tween-20)... 

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