Antibody pairs

In conclusion, based on the literature and our data, the traditional use of acetone-fixed frozen tissue sections as the gold standard for IHC is not justified for all antigen/antibody pairs. For validating any new antibody, it would be prudent to employ a combination of both acetone and NBF-fixed frozen sections. From our experience, FFPE tissue sections may serve as the standard for most antigens for IHC. 

In conclusion, the core components that constitute the mechanism of AR are beginning to come into focus. Further work will be required to solidify these core components and to understand the variations that arise for specific tissues and antigen-antibody pairs. This knowledge, in turn, will lead to improved AR methods, improved ways to overcome the variable effects of tissue fixation, and improved standardization between clinical laboratories. 

After incubation, the supernatants of the ceU cultures can be used for the cytokine assay by capture enzyme-linked immunosorbent assay (ELISA) using specific antibody pairs and recombinant cytokine standards and ELISA plates. 

The titer of the antisera is critical and must be determined experimentally for each antigen-antibody pair on each automated stainer. The ideal titer may... 

Recent laser-tweezer experiments used antibody pairs to demarcate tandem Ig segments within single titin molecules (Leake et al, 2004)... 

Key Words ELISA competition binding substrate antibody pairs standard curve. 

The simplest method for determination of cytokines by ELISA is to use commercially available kits with matching antibody pairs and standards for the specific cytokine. The reader can consult web sites such as http //www.biocompare.com/ to select from a wide range of commercially available kits with the corresponding manufactures instructions for use. 

Lower throughput and wash steps Waddleton (2002) required must have suitable cell Minor (2003) line and antibody pair Zhang (2007)... 

Because of the very large variety of assay detection methods, it is difficult to cover all the parameters needed to optimize for each detection system however, it is important to mention that all the systems discussed in the assay readout technology section above must be optimized. For example, when using an antibody pair in a TR-FRET assay, it is important to find a concentration of antibodies that can trap the product efficiently at the desired time point in the reaction. 

More recently, noncompetitive methods have been developed for measuring human OC. These immunoassays use monoclonal antibodies and affinity-purified polyclonal antisera against synthetic peptides of human OC or intact human or bovine OC. Many antibody pairs have been reported including capture [and signal] antibodies, respectively, against amino acid sequences 12-33 [and 34-49], 43-49 [and 5-13], 25-37 [and 5-13], 20-43 [and 7-19], 1-19 [and 20-49], and 1-12 [and 15-30 or 38-49], These noncompetitive methods have used signal antibodies labeled with radioactivity (IRMA), enzymes (EIA/ELISA), or chemiluminescent compounds (ICMA). 

In sandwich LBAs, both polyclonal antibodies (PAbs) and monoclonal antibodies (MAbs) are used as capture detector antibody pairs in various combinations. As MAbs recognize only a single epitope, they provide higher specificity than PAbs and are consequently more expensive to produce. Conversely, immunization of a single goat or sheep may provide a supply of PAbs sufficient for the execution of multiple studies over several years. PAbs can also provide higher sensitivity due to the presence of multiple antibodies binding to different epitope on the analyte molecule. 

Mossner, E., Koch, H., Pliickthun, A. Fast selection of antibodies without antigen purification adaption of the protein fragment complementation assay to select antigen-antibody pairs./. Mol. Biol. 2001, 308, 115-122. 

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