Preparation Of Specific Antisera Research Articles

Utilização da serologia na identificação de <em>Ascia monuste orseis</em> (Latreille, 1819) (Lepidoptera: Pieridae)

This study aimed to obtain specific antiserum for Ascia monuste orseis (Latreille, 1819), with the aim of identifying the predators associated with this insect. The preparation of specific antiserum was performed by immunizing a rabbit via a lymph nodule injection containing antigen obtained from the maceration of 4 th and 5 th instars of A. monuste orseis caterpillars. Serological homologous tests performed using double diffusion in agar gel were positive 21 days after the first antigen inoculation. Eggs, 1 th , 2 th and 3 th instar caterpillars, pupae and adults of A. monuste orseis also reacted positively to the obtained antiserum.

Welfare of animals kept for harvesting of blood

In New Zealand, blood is harvested from horses, cattle, sheep and goats for research (eg preparation of specific antisera) and commercial purposes (eg vaccine production). Due to the country's favourable disease status there is a growing demand from other countries for harvested blood and it is expected that the blood-harvesting industry in New Zealand will grow. These guidelines were drawn up for use by local Animal Ethics Committees in New Zealand in order to safeguard the welfare of animals used for blood-harvesting. The guidelines contain sections on facilities and equipment, bleeding techniques, volume and frequency of bleeding, animal monitoring (including monitoring of packed cell volume) and other aspects of blood-harvesting. It is stated that not more than 15 per cent of estimated circulating blood volume should be removed in any four week period. However, it may be acceptable in some circumstances, with the approval of the institutional Animal Ethics Committee, for up to 25 per cent of estimated circulating blood volume to be removed. The document is required reading for people involved with the blood-harvesting industry in New Zealand but it is also likely to be of interest and value to those involved in similar work elsewhere in the world.

Penner serotyping of Campylobacter isolates from poultry, with absorbed pooled antisera.

The Penner serotyping system, based on detection of heat-stable antigens with a passive haemagglutination technique, was used in studies on Campylobacter epidemiology in poultry. Preparation of specific antisera by absorption allowed the use of pooled antisera. Over 80% of the Campylobacter isolates were typable with this modified Penner serotyping system. Typability of strains was clearly affected by storage of the strains before actual typing. Extracted antigens appeared to be stable for at least 6 months at 4 degrees C. Therefore, it is advisable to store extracted antigens from freshly isolated Campylobacter strains instead of reculturing frozen-stored strains, when actual typing cannot be performed directly after primary isolation. Untypability of isolates may partly be explained by the detection of Campylobacter serovars not yet represented in the serotyping system. Experiments on repeated serotyping of several Campylobacter strains did not suggest any serovar instability within the strains.

Preparation of specific antiserum to digitoxin by using digitoxin hemisuccinate-bovine serum albumin conjugate at the digitoxose C-3' position

Preparation of specific antiserum to digitoxin by using digitoxin hemisuccinate-bovine serum albumin conjugate at the digitoxose C-3' position

Human intestinal lactase-phlorizin hydrolase: Isolation and preparation of a specific antiserum

Human intestinal lactase-phlorizin hydrolase (lactase) was selectively isolated with monospecific polyclonal antibodies to rat lactase. In addition to their immunologic similarities indicated by this isolation, human and rat lactase have similar kinetic characteristics but different subunit structure when analyzed by gel electrophoresis under reducing conditions. Rabbits immunized by injecting human lactase complexed with anti-rat lactase produced specific antibodies to human lactase that exhibited little cross-reactivity to the rat enzyme. The simple single-step procedure allows isolation of human lactase in high purity from small biologic samples and preparation of specific antisera to the human enzyme.

Preparation of specific antiserum against Vibrio vulnificus hemolysin by immunization with hemolysin-bound liposomes

Specific antiserum against Vibrio vulnificus hemolysin was prepared by immunizing rabbits with a liposome suspension bearing the hemolysin. A cholesterol-phosphatidylcholine-liposome suspension was incubated with crude hemolysin preparation, washed well and then injected intravenously (i.v.) into rabbits. Of the various proteins in the crude preparation, the hemolysin was the only protein which was able to bind to the liposomes, because V. vulnificus hemolysin possesses specific cholesterol-binding activity. Therefore, the antiserum obtained could react only with the hemolysin. Specific antiserum was thus prepared by a simple method, with no need for tedious treatment to purify the immunizing antigen.

Preparation of specific antiserum against Vibrio vulnificus hemolysin by immunization with hemolysin-bound liposomes

Preparation of specific antiserum against Vibrio vulnificus hemolysin by immunization with hemolysin-bound liposomes

A rapid and simple method for preparation of adenovirus DNA from infected cells.

Microbiology and ImmunologyVolume 27, Issue 9 p. 817-822 NoteFree Access A Rapid and Simple Method for Preparation of Adenovirus DNA from Infected Cells Morikazu Shinagawa, Morikazu Shinagawa Department of Veterinary Public Health, School of Veterinary Medicine, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido, 080Search for more papers by this authorAkihisa Matsuda, Akihisa Matsuda Department of Veterinary Public Health, School of Veterinary Medicine, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido, 080Search for more papers by this authorToshiro Ishiyama, Toshiro Ishiyama Department of Veterinary Public Health, School of Veterinary Medicine, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido, 080Search for more papers by this authorHitoshi Goto, Hitoshi Goto Department of Veterinary Microbiology, School of Veterinary Medicine, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido, 080Search for more papers by this authorGihei Sato, Gihei Sato Department of Veterinary Public Health, School of Veterinary Medicine, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido, 080Search for more papers by this author Morikazu Shinagawa, Morikazu Shinagawa Department of Veterinary Public Health, School of Veterinary Medicine, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido, 080Search for more papers by this authorAkihisa Matsuda, Akihisa Matsuda Department of Veterinary Public Health, School of Veterinary Medicine, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido, 080Search for more papers by this authorToshiro Ishiyama, Toshiro Ishiyama Department of Veterinary Public Health, School of Veterinary Medicine, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido, 080Search for more papers by this authorHitoshi Goto, Hitoshi Goto Department of Veterinary Microbiology, School of Veterinary Medicine, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido, 080Search for more papers by this authorGihei Sato, Gihei Sato Department of Veterinary Public Health, School of Veterinary Medicine, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido, 080Search for more papers by this author First published: September 1983 https://doi.org/10.1111/j.1348-0421.1983.tb00638.xCitations: 92AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL REFERENCES 1Carusi, E.A. 1977. Evidence for blocked 5′ termini in human adenovirus DNA. Virology 76: 380– 395. 2Green, M., and Pina, M. 1964. Biochemical studies on adenovirus multiplication, VI. Properties of highly purified tumorigenic human adenoviruses and their DNA's. Proc. Natl. Acad. Sci. U.S.A. 51: 1251– 1259. 3Hirt, B. 1967. Selective extraction of polyoma DNA from infected mouse cell cultures. J. Mol. Biol. 26: 365– 369. 4Horwitz, S.M. 1976. Bidirectional replication of adenovirus type 2 DNA. J. Virol. 18: 307– 315. 5Johansson, M.E., Wadell, G., Jacobson, P.A., and Svensson, L. 1979. Preparation of specific antisera against adenoviruses by affinity bead immunization. J. Immunol. Methods 26: 141– 149. 6Jones, N., and Shenk, T. 1978. Isolation of deletion and substitution mutants of adenovirus type 5. Cell 13: 181– 188. 7Maxam, A.M., and Gilbert, W. 1977. A new method for sequencing DNA. Proc. Natl. Acad. Sci. U.S.A. 74: 560– 564. 8Norrby, E., Bartha, A., Rowe, P., Russell, W.C., Schlesinger, R.W., and Wigand, R. 1976. Adenoviridae. Intervirology 7: 117– 125. 9Padmanabhan, R., and Padmanabhan, R.V. 1977. Specific interaction of protein(s) at or near the termini of adenovirus 2 DNA. Biochem. Biophys. Res. Commun. 75: 955– 964. 10Pettersson, U., and Sambrook, J. 1973. Amount of viral DNA in the genome of cells transformed by adenovirus type 2. J. Mol. Biol. 73: 125– 130. 11Rekosh, D.M.K., Russel, W.C., Bellett, A.J.D., and Robinson, A.J. 1977. Identification of a protein linked to the ends of adenovirus DNA. Cell 11: 283– 295. 12Robinson, A.J., Younghusband, H.B., and Bellett, A.J.D. 1973. A circular DNA-protein complex from adenoviruses. Virology 56: 54– 69. 13Sharp, P.A., Moor, C., and Haverty, J.L. 1976. The infectivity of adenovirus 5 DNA-protein complex. Virology 75: 442– 456. 14Shinagawa, M., and Padmanabhan, R. 1979. Inhibition of a nuclease contamination in the commercial preparations of Escherichia coli alkaline phosphatase. Anal. Biochem. 95: 458– 464. 15Shinagawa, M., and Padmanabhan, R. 1980. Comparative sequence analysis of the inverted terminal repetitions from different adenoviruses. Proc. Natl. Acad. Sci. U.S.A. 77: 3831– 3835. 16Wimmer, E. 1982. Genome-linked proteins of viruses. Cell 28: 3831– 3835. Citing Literature Volume27, Issue9September 1983Pages 817-822 ReferencesRelatedInformation

Preparation of Specific Antisera to Honeybee Viruses by Immunization with Agar Gel Precipitates

SummaryA technique for the production of specific antisera to sac brood virus (SBV), chronic bee paralysis virus(CBPV) and Kashmir bee virus (KBV) is described. Antisera suitable for gel-diffusion precipitin tests were prepared against SBV, CBPV and KBV by immunizing rabbits with specific agar gel immune precipitates. These precipitates were produced in gel-diffusion precipitin tests using partially purified viral extracts and reference antisera.

Human T cell leukemia antigens on the cell membranes: purification molecular characterization, and preparation of specific antisera.

Human T cell leukemia antigens on the cell membranes: purification molecular characterization, and preparation of specific antisera.

Preparation of specific antisera against adenoviruses by affinity bead immunization (ABI)

Certain adenovirus types can be replicated only to low titer in tissue cultures. Other, such as adenovirus strains associated with infantile gastroenteritis, cannot be replicated in vitro. A method which allows preparation of specific antisera has therefore been evaluated. The procedure involves coupling of group-specific antibodies against adenovirus capsid subunits to CNBr-activated Sepharose 4B; reaction of crude virus suspensions with immobilized adenovirus-specific IgG; elimination of contaminating material by extensive washing using a wide pH range; and immunization with adenovirus immunogens immobilized on the beads. Efficient immunization was obtained with immunogen doses of both 50 ng and 50 μg. The immunization procedure which has been designated affinity bead immunization (ABI) could therefore have a wide applicability in cases where the relevant immunogen constitutes a minor fraction of a crude preparation.

Steroid Hormone Action: Recent Advances

Pn entering the cell, steroid hormones are bound to specific cytoplasmic receptors. The hormone-receptor complexes are then translocated to the nucleus, in an "activated" form, whereupon they are bound to the target cell genome. The target cell responds by increased RNA synthesis with the transcription of specific mRNAs. The mRNAs are exported to the cytoplasm where protein synthesis takes place. Recent advances in steroid hormone action involve the purification of specific steroid hormone receptors and the preparation of specific antisera, the elucidation of the subunit structure of the progesterone receptor, the purification of a hormone-inducible mRNA and the synthesis of its DNA complement, the purification and amplification of the synthetic and natural gene for this RNA and the identification of intragenic spacers, and most recently the identification of specific precursors to the hormone-induced mRNA. We discuss here the medical significance of some of these advances.

Glutamine synthetase from rat liver. Purification, properties, and preparation of specific antisera.

Glutamine synthetase from rat liver. Purification, properties, and preparation of specific antisera.

Preparation of specific antisera to Drosophila acid phosphatase without rigorous protein purification.

Extracts from an acid phosphatase CRM- null mutant of Drosophila melanogaster were used to eliminate contaminating antibodies in a nonspecific preparation of anti-acid phosphatase serum. This method of producing specific antisera makes unnecessary the rigorous purification of an antigen prior to immunization attempts in those cases where CRM- null mutants of the antigen are available. Antisera so prepared could be used for a wide variety of purposes.

Immunochemical identification of carcinoembryonic antigen in ovarian adenocarcinoma and pseudomucinous cystoma extracts

Carcinoembryonic antigen (CEA) was found in 3 of 17 individual extracts of adenocarcinoma and in 7 or 8 individual samples of pseudomucinous cystoma of the human ovaries. Rabbits were immunized for the purpose of preparation of specific antisera against CEA by individual fractions (ammonium sulfate, sulfosalicylic acid and phosphotungstic acid) of CEA-positive adenocarcinomas of the ovary.

A Specific Precipitin Antiserum for Carcinoma Protein

Attempts have been made by numerous investigators to prepare antiserum for normal and malignant tissue protein (1). These antisera necessarily contained antibodies for blood serum proteins. No matter how finely tissue is ground and how thoroughly it is washed, it always contains sufficient blood proteins to elicit antibodies. Lumsden (2) believed that he could wash ground tissue free from blood, but the toxicity of his antiserum may have been due to antibodies for serum proteins. When the precipitins for blood serum proteins are removed from antiserum by adsorption with their antigens, antibodies for tissue proteins are also partially or completely removed. Spinka and Weichselbaum (3) were the first to attempt the preparation of specific antiserum for kidney and liver proteins by fixing the finely ground washed tissue on aluminium cream and injecting this material intramuscularly into rabbits. Their attempts to remove the blood serum protein precipitins from the antiserum by adsorption resulted in the removal of all or practically all tissue precipitins. They were not successful in their attempt to apply the adsorption in vivo method (4). They discovered, however, that rabbits injected with tissue proteins, after a period of months, lost the blood serum precipitins, whereas the precipitins for tissue proteins remained in relatively high titers. Hektoen and Welker (5) previously observed that rabbits, immunized with aluminium cream on which many antigens had been adsorbed, lost the precipitins for the different antigens at different intervals. This disappearance seemed to bear a relation to the quantity of the different antigens in the injected cream. It is to be expected that as a given antigen is exhausted from the aluminium cream depot, the related precipitin antibody will sooner or later disappear from the blood. We have applied this method in the case of certain antigens with traces of other antigens. On injection, precipitins will be called forth for all the antigens, but the precipitins for traces of antigens soon disappear.