Antiserum

The double-immunodiffusion technique, often referred to as the Ouchtedony technique, uses an agarose gel as the matrix. Holes are made in the agarose where either sample or antisera is placed. The two solutions are allowed to diffuse into the matrix for a predetermined time. If there is a reaction... 

By 1945, Stacey speculated about the possibility of a structural relationship between Pneumococcus capsular polysaccharides and those produced by other organisms. With Miss Schliichterer, he had examined the capsular polysaccharide of Rhizobium radicicolum. This polysaccharide gave a precipitin reaction in high dilution, not only with Type III Pneumococcus antiserum, but also mixed with antisera from other Pneumococcus types. The chemical evidence indicated that the polysaccharide resembled the specific polysaccharides of Types I and II Pneumococcus. A decade later, the acidic capsular polysaccharide from Azoto-bacter chroococcum, a soil organism, was studied. It, too, produced serological cross-reactions with certain pneumococcal specific antisera. Although the molecular structure of the polysaccharide was not established, adequate evidence was accumulated to show a structural relationship to Type III Pneumococcus-specific polysaccharide. This was sufficiently close to account for the Type III serological cross-relationship. 

An alternative pathway for activating the cascade has recently been demonstrated in which factor XII is absent from the reaction mixture [42-45]. Two different groups have isolated two different proteins, each of which seems to activate the HK-prekallikrein complex. One is heat-shock protein 90 [46] and the other is a prolylcarboxypeptidase [47]. Neither protein is a direct prekallikrein activator as is factor Xlla or factor Xllf because each activator requires HK to be complexed to the prekallikrein. In addition, the reaction is stoichiometric, thus the amount of prekallikrein converted to kallikrein equals the molar input of heat-shock protein 90 (or prolylcarboxypeptidase). These proteins can be shown to contribute to factor Xll-independent prekallikrein activation and antisera to each protein have been shown to inhibit the process. When whole endothelial cells are incubated with normal plasma or factor Xll-deficient plasma, the rate of activation of the deficient plasma is very much slower than that of the normal plasma, the latter being factor Xll-dependent [45]. Under normal circumstances (with factor XII present), formation of any kallikrein will lead to factor Xlla formation even if the process were initiated by one of these cell-derived factors. 

Synthetic hCT Is not commercially available now. Most assays have been developed by using hormone supplied as gifts from the CIBA-Gelgy Co., Basel, Switzerland for 131i or labeling. Immunization and standards although several useful antisera have been produced by Immunization with crude extracts of medullary carcinoma tissue. 

Inoculation of cell cultures with virus-containing material produces characteristic changes in the cells. The replication of many types of viruses produces the cytopathic effect (CPE) in which cells degenerate. This effect is seen as the shrinkage or sometimes ballooning of cells and the disruption of the monolayer by death and detachment of the cells (Fig. 3.6). The replicating virus can then be identified by inoculating a series of cell cultures with mixtures of the virus and different known viral antisera. If the virus is the same as one of the types used to prepare the various antisera, then its activity will be neutralized by that particular antiserum and CPE will not be apparent in that tube. Alternatively viral antisera labelled with a fluorescent dye can be used to identify the virus in the cell culture. 

Wall, SJ, Yasada, RP, Hory, F, Flagg, S, Martin, BM, Ginns, El and Wolfe, BB (1991) Production of antisera selective for Mi muscarinic receptors using fusion proteins distribution of Ml receptors in rat brain. Mol. Pharmacol. 39 643-649. 

Polyclonal Antibodies against FORL r purified polygalacturonase were raised in white rabbits. For the first immunization 200 jxg of purified protein in 300 jxl of distilled water was mixed with 200 1 of PBS and 500 n of complete Freund s adjuvcmt and injected intramusculary into the leg. Two subsequent intramuscular injections, each containing 300 fig of protein in 1 ml of incomplete Freund s adjuvant were given at 1 month intervals. Finally, the rabbit was bled 1 week later. The antisera, separated from blood by incubation at 37 "C, were stored in 1 ml fractions at -20 C. 

Specificity of the antisera was assessed by Western blotting. Electrophoretically separated proteins from culture filtrates were transferred to 0.45 fim nitrocellulose membranes. After transfer of proteins, membranes were... 

In a preliminary report, Ross et al. [40] used affinity chromatography to identify a putative bovine renal brush border Na /H exchanger. Brush border membranes were solubilized with Triton X-100 and chromatographed sequentially over lentil lectin Sepharose 4B and 5-(A-benzyl-iV-ethyl)amiloride coupled to epoxy-activated Sepharose 6B. The eluant contained 178- and 146-kDa proteins that were susceptible to Endo-F. Moreover, the eluants reacted on dot blot immunoassays with antisera to a 20-amino acid peptide of a human Na /H exchanger vide infra). The relationship between these proteins and the 66-kDa protein previously identified by the same investigators using amiloride photolabeling is presently unclear. 

In contrast to these results, Ross et al. [33] found that antisera against a 20-amino acid peptide (Ser-613-Arg-632) of the cytoplasmic domain of the human Na /H exchanger recognized a 66-kDa protein in immunoblots of bovine renal brush border membranes. Since the purity of these membranes was not reported it is possible that this result was due to contamination with basolateral membranes (although the molecular mass would still differ from the basolateral Na /H exchanger in LLC-... 

Palinski, W., Yla-Herttuala, S., Rosenfeld, M.E., Butler, S.W., Socher, S.A., Parthasarathy, S., Curtiss, L.K. and Witztum, J.L. (1990). Antisera and monoclonal antibodies specific for epitopes generated during oxidative modification of low density lipoproteins. Arteriosclerosis 10, 325-335. 

EHEC belonging to serotype 0157 H7 characteristically do not ferment sorbitol, whereas more than 70% of intestinal floral f. coli do. To properly screen EHEC strains in cases of diarrhea, stool should be placed on special sorbitol-MacConkey agar. Colonies of f. coli 0157 H7, which do not ferment the sorbitol, can be identified readily and confirmed by serotyping with specific antisera. In addition, stool should be tested directly for the presence of Stx I and II by enzyme immunoassay (EIA). 

Gruszecki, W.I., A. Sujak, K. Strzalka, A. Radunz, and G.H. Schmid. 1999. Organisation of xanthophyll-lipid membranes studied by means of specific pigment antisera, spectrophotometry and monomolecular layer technique lutein versus zeaxanthin. Z. Naturforsch. C 54 517-525. 

Abramowski, D., Rigo, M., Due, D., Hoyer, D. Staufenbiel, M. (1995). Localization of the 5-hydroxytryptamine2c receptor protein in human and rat brain using specific antisera. Neuropharmacology 34, 1635-45. 

Szewczyk B, Summers DF (1987) Fluorescent staining of proteins transferred to nitrocellulose allowing for subsequent probing with antisera. Anal Biochem 164 303-306... 

Fig. 14.4. Expression of subunits of the pyruvate dehydrogenase complex during the development of A suum. Homogenates of different A suum larval stages and adult tissues were immunoblotted with polyclonal antisera prepared against individual subunits of the A suum PDC isolated from adult muscle, as described in detail in Klingbeil etal. (1996). UE, unembryonated egg M, adult body wall muscle p45, E3-binding protein (E3BP).

Bland, P.W. and Warren, L.G. (1986) Antigen presentation by epithelial cells of the rat small intestine. I. Kinetics, antigen specificity and blocking by anti-la antisera. Immunology 58, 1-7. 

Although immunocytochemical screening of neurochemical diversity provides only an indication that a neuropeptide may be present, antisera that interact specifically with known nematode neuropeptides are likely... 

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