Buffers coating phosphate-buffered saline

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,... 

Freshly excised buccal or sublingual tissues have also been used to generate dissociated cells. Hedberg et al. [49] used one such culture to measure the expression of alcohol dehydrogenase-3 in cultured cells from human oral mucosal tissue. Human buccal tissue was incubated with 0.17% trypsin in phosphate-buffered saline (PBS) at 4°C for 18 to 24 h to obtain dissociated primary keratinocytes, and subsequently these keratinocytes were seeded onto fibronectin and collagen-coated dishes in serum-free epithelial medium. 

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.

Fig. I. Crosman BB shot beads plated with bright nickel and black nickel (for color coding purposes), then coated with polycarbonate by immersing in 5% solution of methylene chloride, scattering on slick paper, and allowing to dry. Various antigens or antibodies are adsorbed onto this plastic surface the beads are soaked in a % solution of bovine serum albumin in phosphate-buffered saline, drained, air dried, and stored below 0° until needed.

Fig. 10.2. Comparison of optical and hydrodynamic properties of CdTe quantum dots (2.5 nm) solubilized in water with an amphiphilic polymer (octylamine-modified polyacrylic acid) or a multidentate polymer ligand, (a) Absorption (blue curves) and fluorescence emission red curves) spectra of CdTe quantum dots with amphiphilic polymer upper) or multidentate polymer lower) coatings, (b) Dynamic light scattering size data of quantum dots with amphiphilic polymer blue curve) and multidentate polymer green curve) coatings. PL Photoluminescence, AU Arbitrary units. All samples were dissolved in phosphate buffered saline...

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... 

The conjugate of biotinylation-anti-albumin was synthesized and purified as described in the paper. Biotinamidohexanoyl-6-aminohexanoic acid N-hydroxysuccinimide ester (BCNHS) with an extended spacer arm was applied to react with primary amines of the protein which can reduce the steric hindrance. Avidin-biotin chemiluminescence immunoassay procedure Immunoassay procedures are presented in Figure. 1. The wells of the microtiter plates were coated with 150 pL of avidin (3 mg/mL) in a phosphate buffered saline... 

To develop a highly consistent solid phase, coating and blocking buffer and temperature were studied. It was found that the optimal coating buffer was 10 mM phosphate-buffered saline (PBS), pH 7.0. 

Cook s microtiter wells are coated with 50 pL of 10 pg/mL solution of human cardiac myosin, then the plates are blocked and washed 12 times with 1% horse serum in 0.15 M phosphate-buffered saline (pH 7.4). 

We also reported a concept for a sandwich immunoassay that is completely performed on-chip using streptavidin-coated beads as substrate [17]. The latter were electrostatically self-assembled on aminosilane micropattems at the bottom of a microfluidic channel. We used mouse IgG diluted in phosphate buffered saline (PBS) with 1% bovine serum albumin (BSA) solution as target antigen. The fluorescent sandwich immunocomplex was formed on the beads during the operation of the chip both in stop-flow and continuous flow modes. Target mouse IgG antigen could be detected down to a concentration of 15 ng/mL in stop-flow mode and 250 pg/mL in continuous flow mode, using only 1,300 nL of sample volume. 

Freezer -80°C for storing tissue Cryostat with several chucks Coated microscope slides Phosphate buffered saline (PBS)... 

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