Plate polystyrene microtiter

Preparation of microtiter plates. A constant amount of the coating antigen is bound to the surface of polystyrene microtiter plate wells by passive adsorption. After a predetermined incubation time, the plate is washed to remove unbound coating antigen. 

Figure 3 Immobilized antibody ELISA. Primary antibody (Y) is passively adsorbed to the surface of a polystyrene microtiter plate. Analyte (free H) and an enzyme-labeled hapten (H-E) compete for the fixed number of primary antibody binding sites. Following a wash step (dotted line), the substrate for the enzyme is added O) and a colored product formed ( ). The amount of product is inversely proportional to the amount of analyte present...

Other early work includes that of Moody et al. (2001) who spotted anticytokine monoclonals onto the bottom of polystyrene microtiter plates (Max-isorp, Nalge Nunc) and measured cytokine levels in stimulated peripheral blood mononuclear cells. Finally, although not strictly a microarray, the microwell array system developed by Michael Snyder s group at Yale University to measure kinase activity is a simple and elegant approach (Zhu et al., 2000). The "protein chip" is comprised of microwells fabricated in a flexible elastomer of PDMS [poly(dimethylsiloxane)] substrate by a molding process. 

Enzyme-Immunoassay. Fish tissue samples for testing were cut into uniform 3mm thick slices with parallel razor blades mounted on a handle. Four discs were then punched out from each slice with a stainless steel borer, 3-mm in diameter, and each disc was placed in a well of a 96-well polystyrene microtiter plate (Flow Laboratories, Inc., Hamden, CT). Samples were washed once with 0.2 ml Tris buffer. After the wash solution was aspirated, each sample was fixed in 0.2 ml of 0.3% H O -methanol fixative for 30 min. at room temperature. Samples were then transferred to clean wells and 0.2 ml of a 1 100 dilution of sheep-anti-ciguatoxin-horseradish peroxidase conjugate in Tris buffer was added to each well. The plate was then incubated at room temperature for 1 hr. The sheep-anti-cigua-toxin-horseradish peroxidase was removed by aspiration, and the tissues were immersed for 5 min. in 0.2 ml Tris buffer. Each sample was transferred to clean wells and incubated for 5 min. at room temperature with 0.2 ml of 4-chloro-l-naphthol substrate. The final steps involved removal of the tissue and addition of 0.015 ml of 3 M sodium hydroxide to stop the enzymatic reaction. Absorbance readings at 405 nm of each well were obtained in the Titertek Multi-skan (Flow Laboratories, Inc., Hamden, CT). 

Figure 8 Chemiluminescent (A and B) and bioluminescent (C) detections for immobilized hybridizations of PCR product. Dg, digoxigenin Bt, biotin Ad, avidin. Procedure A [30] Biotin moiety is incorporated into PCR products during the amplification reaction, using one 5 -biotinylated primer. The product is hybridized with a Dg-labeled probe and is immobilized on streptavidin-coated magnetic beads. This capture reaction is carried out for 30 min at 37°C. A permanent magnet is used to sediment the beads during washing to remove unbound DNA. By incubation with the washed beads for 45 min at 37°C, anti-Dg antibody conjugated to HRP enzyme is bound to the Dg-labeled probe, and luminol reaction is performed for CL detection. Procedure B [31] Wells of a polystyrene microtiter plate are activated with l-ethyl-3-(3-dimethylaminopropyl)-carbodiimide, and then coated with a labeled cDNA probe complementary to an internal region of the target DNA.

The ELISA procedure for the analysis of parathion as described above requires nearly eight hours, although many samples can be simultaneously assayed. However, incubation times can be shortened to one-half hour, in most cases, resulting in only a 10% reduction in sensitivity. Also the polystyrene microtiter plates containing bound RSA-AP can be mass produced and stored in a freezer. Since the enzyme-linked antibody can be purchased, the limiting factor of the applicability of the ELISA procedure, as well as the RIA procedures, for other pesticides is the development of the antiserum to the pesticide. 

Polystyrene microtiter plates are coated with purified target, washed to remove the excess of the protein, then saturated with a specific reagent, such as BSA, dried milk, or gelatin, to block the uncoated plastic surface. 

Many assays that have been bead-captured and gel-based have been converted to 96-well polystyrene microtiter plates to standardize the format for automation and higher throughput. In many ways, plate formats are a bridge to the higher-density microarray formats and remain quite useful until full conversion to microarray or microfluidic formats can be made. Microtiter plate assays or activated microbeads (i.e., Luminex) for panels of cytokines are widely available because of their use in clinical medicine. 

Enzymes are currently the most widely used and investigated labels for immunoassays, because a single enzyme label can provide multiple copies of detectable species. This catalytic amplification results in immunoassay detection limits that rival those of radioimmunoassay without the storage and disposal problems associated with radioisotopes. Enzyme immunoassays label either ligands or antibodies with enzyme, and enzyme activity in bound or free fractions is measured. Heterogeneous immunoassays employing enzymatic labels have been named enzyme-linked immunosorbent assays (ELISAs). ELISA methods usually employ antibody immobilized onto the wells of polystyrene microtiter plates, and may be... 

Polystyrene microtiter plates are coated with purified target. 

Polystyrene Microtiter plates, tubes, pins, beads Noncovalent 250-500 ngcm ... 

Solutions were prepared from bidistilled water. BSA (fraction V, Boehringer-Mannheim) was used as a standard protein. Ninety-six-well, flat-bottomed polystyrene microtiter plates (Greiner, Cat. No. 655101) were used for the photometric tests. 

This assay is the most important in trace and environmental analysis. First, antibodies are immobilized on a solid carrier. For this, it may be necessary to take suitable measures, such as Co radiation, to make the carrier sorption active. Standard 96-well microtiter plates, polystyrene beads, or test tubes can be used. 

Rodrfguez JC, Pastor E, Ruiz M, Flores E, Royo G (2007) Antibiotic resistance during therapy mechanisms and means of control. Infect Disord Drug Targets 7 43-45 Rukayadi Y, Hwang JK (2006) Effect of coating the weUs of a polystyrene microtiter plate with xanthorrhizol on the biofilm formation of Streptococcus mutans. J Basic Microbiol 46 410- 15... 

For higher throughput applications, injection-molded plastic microtiter plates have served as the formats of choice for automated assay development. Thermoplastics such as polystyrene, polycarbonate, and polypropylene are used for a variety of purposes including storage and assay plates, lids, pipette tips, and Eppendorf PCR tubes. Polystyrene plates are used for cell culture and ELISAs. Polycarbonate reagent bottles are popular, while polypropylene storage plates and PCR tubes are standards. 

The ELISA procedure recently has been used for the analysis of parathion (31). Since this procedure has considerable potential a more detailed description of the analysis of parathion is in order. The conjugation procedure using amino parathion (AP) was described earlier (Fig. 3, Rn 9), and this conjugate was then administered to rabbits for development of a population of specific antibodies (Abi) against BSA or AP. Abi demonstrated immunological activity only for the hapten when AP was conjugated to rabbit serum albumin (RSA). This antigen (RSA-AP) was rendered insoluble via attachment to the polystyrene surface of microtiter plates under basic conditions (Fig. 6.1). 

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