• Title/Summary/Keyword: Antisera

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Production of Standard Antisera for the Blood Typing in Cattle (동종면역에 의한 소 혈액형 표준혈청의 생산)

  • 신형두;한호재;이국경;강동묵;양일석;권종국
    • Journal of Embryo Transfer
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    • v.3 no.1
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    • pp.31-37
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    • 1988
  • This study was carried out to produce the antisera for the blood typing in cattle. Blood types of eighty cattle were previously determined by 56 kinds of internationally standardized antisera from Japan. The donorrecipient animal arrangements were determined according to tile previously determined blood types of animals by the computer program SS-l for efficient production of antisera. Six kinds of standard antisera, H,B', 12, C2, Z, U2, were produced by isoimmunirzation and absorption methods.

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Production and identification of antisera against mu-opioid receptor using synthetic peptide epitope (Synthetic peptide를 이용한 mu-opioid receptor에 대한 항혈청의 생산과 검정)

  • Lee, Jang-hern;Kwon, Young-bae;Han, Ho-jae
    • Korean Journal of Veterinary Research
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    • v.39 no.1
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    • pp.45-54
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    • 1999
  • In the present study we have analyzed the characteristics and distribution of the mu-opioid receptor(MOR) by raising anti-peptide antisera to the C-terminal peptide of MOR. The antisera against MOR was produced in New Zealand White rabbit against 15 residue corresponding to amino acids, 384-398 of the cloned rat MOR. The antigenic peptide was synthesized using an Applied Biosystems 432 solid-phase peptide synthesizer. The specificity and identification of the antisera were tested by analysis of transfected cells, epitope mapping and immunohistochemical method. COS-7 cells electroporated with MOR cDNA were used to evaluate the characteristics and subcellular distribution of MOR. MOR immunoreactivity was prodominent in the plasmalemma and subcellular compartments such as endoplasmic reticulum, Golgi apparatus and vesicle like structure. Furthermore, both tissue sections and transfected cell lines could be immunostained with these antisera and the immunoreactivity was abolished when anti-MOR sera were preincubated with the peptide against which they were raised. Based on epitope mapping analysis, all antisera appeared to have a similar epitope, which was determined to be within the last amino acid, 391-398. Moreover, immunohistochemistry showed that MOR immunoreactivity was observed in many brain areas including cerebral cortex, striatum, hippocampus, locus coeruleus and the superficial laminae of the dorsal horn. These stained spinal cord and brain areas showed the mirrored pattern observed in auto radiographic studies of mu-opioid binding as well as a pattern similar to that seen by is situ hybridization for MOR. Thus, several lines of evidence support the conclusion that the antisera produced in the present study most likely recognize mu-opioid receptor. These results suggest that MOR antisera may be utilized as useful tool to analyze the physiological and pharmacological studies for mu-opioid receptor in the future.

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Immunological Variations of Flagella Antigens in Bacillus thuringiensis serovar kurstaki Temperature-sensitive Mutants (Bacillus thuringiensis serovar kurstaki 돌연변이 균주의 편모항원의 변이성)

  • 이형환;이영주;이희무
    • Microbiology and Biotechnology Letters
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    • v.17 no.1
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    • pp.81-83
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    • 1989
  • The flagella antigenic variation of nine Bacillus thuringiensis serovar kurstaki temperature-sensitive mutants grown at the permissive temperature (3$0^{\circ}C$) was detected by a serological agglutination between H-antigen and antiserum. The flagella antigens were injected to rabbits to prepared their antisera, and then their homologous and heterologous titers of the antisera were measured. The homologous titers were ranged from 1:6,400 to 1:12,800, but the heterologous titers were very low. The H-antigen of the wild type strain was not agglutinated to 4 heterologous antisera, ts-U23 not to 7, ts-U3l not 5, ts-U32 not to 4, ts-U33 not to 7, ts-U7l not to 4, ts-U73 not to 6, ts-U74 not to 6, ts-U91 not to 4 and ts-U603 not to 4 antisera.

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Inhibition of Yeast Growth by Broadly Cross-Reactive Antisera Elicited by Heterologous Mannan-Protein Conjugate

  • Machova, Eva;Korcova, Jana;Cizova, Alzbeta;Bystricky, Slavomir
    • Journal of Microbiology and Biotechnology
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    • v.25 no.7
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    • pp.1177-1179
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    • 2015
  • A new approach to obtain broadly cross-reactive antisera against important yeast pathogens by intensive hyperimmunization with polysaccharide-protein conjugates is described here. Surface mannan of Candida albicans and capsular galactoglucoxylomannan of Cryptococcus laurentii were isolated and chemically linked to human serum albumin. Antisera elicited by a 7-week vigorous immunization of rabbits with the conjugates showed effective cross-reactive growth inhibition of different representatives of Candida spp. as well as Cryptococcus spp. IgG antibodies are evidenced as the effective component of the antisera.

A Systematic Study on Selected Amentiferous Plant Taxa - By Qualitative Analysis of Proteins - (유이화서 식물군의 계통분류학적 연구 - 단백질의 정성분석적 접근 -)

  • 이유성
    • Journal of Plant Biology
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    • v.28 no.3
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    • pp.179-190
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    • 1985
  • A review of taxonomic treatments of the amentiferous plant taxa reveals diverse classification. It appears to be necessary to investigate new characteristics and attempt to determine the significance of these characteristics in terms of amentiferous systematics. Serological techniques have been empolyed in a systematic investigation of selected taxa of the Amentiferae: Betulaceae, Fagaceae, Juglandaceae, Myricaceae, and Salicaceae. Data by qualitative analysis of pollen proteins, double immunodiffusion and conventional immunoelectrophoresis, have proved to be valuable in delimitation of taxa tested. When the antisera against Alnus hirsuta, Betula platyphylla var. japonica, Quercus aliena, Q. dentata, Q. glauca were tested; Alnus and Betula have the greatest protein similarity to one another; and next to the rest of betulaceous genera. Relatively strong protein similarity obtained with most representatives of Quercus and Fagus when reacted with antisera against Alnus and Betula is very much in contrast to the weak protein similarity obtained for the genera of Juglandaceae, Myricaceae, particularly of Salicaceae Tested with the same antisera. When Quercus antisera were used for various genera, the weakest protein similarity was obtained with Populus.

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Serological Study on the Cross-Reactivity of Bacteroides gingivalis, Bacteroides intermedius and Bacteroides asaccharolyticus by Indirect Immunofluorescence and Enzyme- Linked Immunosorbent Assay (형광 현미경법 및 효소결합 면역흡착법을 이용한 Bacteroides gingivalis, Bacteroides intermedius 및 Bacteriudes asaccharolyticus의 혈청학적 연구)

  • Chung, C.P.;Lee, J.Y.;Lee, Y.H.;Chung, H.W.;Chung, H.J.
    • The Journal of the Korean Society for Microbiology
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    • v.22 no.2
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    • pp.117-123
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    • 1987
  • Previous studies have been performed for the sero-identification of selected species of Bacteroides by immunofluorescence antibody techniques and enzyme-linked immunosorbent assay using species-specific rabbit antisera to B. gingivalis, B. intermedius, and B. melaninogenicus. However, these studies have not commented on the serological cross-reactivity between these 3 species of black- pigmented Bacteroides. For the cross-reactivity study, antisera to B. gingivalis ATCC 33277, B. intermedius ATC C25261 and B. asaccharolyticus ATCC 25260 were raised from rabbits. Preliminary study for observing the cross-reactivity between these species was performed by indirect immunoflourescence technique. Immunoabsorption of the antisera was done with bacterial cells from the other species and the species-specificity of the antisera was conformed by the absence of reactivity with bacterial strains from the other species by indirect immunofluorescence technique and enzyme-linked immunosorbent assay. Three representative unabsorbed antisera cross-reacted strongly with cells from the other species. Especially, anti-B. asaccharolyticus ATCC 25260 antiserum showed a strong cross-reactivity with B. gingivalis ATCC33277. After immunoabsorption of 3 representative antisera with the other species, the cross-reactivity was found only between B. gingivalis ATCC 33277 and B. asaccharolyticus ATCC 25260. Further study would be necessary to clarify the cross-reactivity between important oral black-pigmented Bacteroides from subgingival plaque or bacterial colonies for rapid identification.

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Production of Polyclonal Antibodies Specific to Porcine Adipocyte Plasma Membrane Proteins in Sheep (면양을 이용한 돼지 지방제포 원형질막 단백질 특이 항체의 생산)

  • 최창본;이명진;권은진
    • Biomedical Science Letters
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    • v.4 no.1
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    • pp.57-63
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    • 1998
  • The objectives of this study were to produce polyclonal antibody to adipocyte plasma membrane (APM) proteins isolated from pig, and to investigate its tissue specificity. Plasma membrane proteins from adipocyte, brain, heart, kidney, liver and spleen were isolated using a self-forming Percoll gradient. Sheep (40kg) was immunized three times at three week interval with the purified APM proteins. Blood was taken from non-immunized sheep (NS) and from immunized sheep at 10 (AS-1), 12 (AS-2), and 14 (AS-3) days after the third immunization. Antisera titers and cross-reactivity against other tissues were determined by enzyme-linked immunosorbent assay (ELISA). Antisera reacted strongly to APM proteins showing detectable amounts of antibody at 1:81,000 dilution. And antisera showed much stronger reactivity to APM proteins than any other tissue plasma membrane proteins. Furthermore, tissue specificity of antisera against APM was reconfirmed by immunoblotting using anti-sheep immunoglobulin G-horseradish peroxidase conjugate as a secondary antibody Antisera to APM proteins showed adipocyte specificity compared with other tissues. In conclusion, polyclonal antibody against APM proteins isolated from pig was developed successfully in our laboratory, and these antisera showed tissue specificity with APM.

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Development of an Indirect ELISA and Immunocapture RT-PCR for Lily Virus Detection

  • Kim, Jin Ha;Yoo, Ha Na;Bae, Eun Hye;Jung, Yong-Tae
    • Journal of Microbiology and Biotechnology
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    • v.22 no.12
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    • pp.1776-1781
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    • 2012
  • Multiple viruses such as Lily symptomless virus (LSV), Lily mottle virus (LMoV), and cucumber mosaic virus (CMV) are the most prevalent viruses infecting lilies in Korea. Leaf samples and bulbs showing characteristic symptoms of virus infection were collected from Gangwon, Chungnam, and Jeju provinces of Korea in 2008-2011. Coat protein (CP) genes of LSV and LMoV were amplified from collected samples by reverse transcription-polymerase chain reaction (RT-PCR) and cloned into a pET21d(+) expression vector to generate recombinant CPs. The resulting carboxy-terminal His-tagged CPs were expressed in Escherichia coli strain BL21(DE3) by isopropyl-1-thio-${\beta}$-D-galactoside induction. The recombinant proteins were purified using Ni-NTA agarose beads, and the purified proteins were used as an immunogen to produce polyclonal antibodies in rabbits. The resulting polyclonal antisera recognized specifically LSV and LMoV from infected plant tissues in Western blotting assays. Indirect enzymelinked immunosorbent assay and immunocapture RT-PCR using these polyclonal antisera were developed for the sensitive, efficient, economic, and rapid detection of Lily viruses. These results suggest that large-scale bulb tests and economic detection of Lily viruses in epidemiological studies can be performed routinely using these polyclonal antisera.

Studies on the Preparation of $T_3-BSA,\;T_4-BSA$ Conjugates, and Radioimmunoassay Use of the Produced Antisera ($T_3-BSA,\;T_4-BSA$ 접합체 제조 및 생성항체의 방사면역측정적 이용 연구)

  • Awh, Ok-Doo;Kim, Jae-Rok
    • The Korean Journal of Nuclear Medicine
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    • v.14 no.1
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    • pp.45-56
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    • 1980
  • $T_3-BSA\;and\;T_4-BSA$ conjugates were prepared and identified spectrophotometrically. The ${\lambda}max$ of the conjugates was just coincided with that of BSA, but the molar extinction coefficients of the conjugates were generally larger than that of BSA itself. The molar ratios of $T_3:\;BSA\;and\;T_4:\;BSA$ in the prepared conjugates were found to be 9:1 and 5:1, respectively. The titers of the $T_3$ antisera were generally higher (max. $1.5{\times}10^4:1$) than those of $T_3$ (max. $2{\times}10^3:1$), and the average cross reactivity of the $T_3$ antibody with $T_3$ was lower (0.45%) than that of $T_4$ antibody with $T_3(3{\sim}4%)$. The results of the study indicate that the predominant cause of the lower titers and the lower specificity of the $T_4$ antisera comparing with those of $T_3$ is mainly due to the unstability of the $T_4-BSA$ and consequent degradation of the conjugate to $T_3-BSA$ during preparation, purification, and even during immunization. The lower molar ratio of $T_4$ to BSA in the preparation stage is also considered to be a minor factor. By measuring $T_3,\;T_4$ levels in the reference control serum, it has been confirmed that the prepared antisera can sufficiently be utilized, respectively, in the established radioimmunoassay systems.

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Expression of VP2 of Aquatic Birnavirus GC-1 Isolated from Rockfish (Sebastes schlegeli), Rearing in Seawater in Korea (국내 해산양식어 조피볼락에서 분리된 수생버나바이러스 GC-1의 VP2 발현)

  • Joh, Seong-joon;Sung, Haan-woo;Lee, Yun-jeong;Kim, Jae-hong;Kang, Shien-young
    • Korean Journal of Veterinary Research
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    • v.43 no.3
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    • pp.449-456
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    • 2003
  • The VP2 gene of aquatic birnavirus, Korean isolate (GC-1) was cloned and expressed using the baculovirus expression system. The VP2 gene and VP2 partial gene, which contained a neutralizing epitope, were constructed for recombinant transfer vectors, for baculovirus expression. The expressed recombinant proteins were confirmed by indirect immuno fluorescence antibody (IFA), SDS-PAGE and Western blot. The level of expression was checked at regular time using IFA and Western blot. To measure the neutralizing activity of recombinant proteins against GC-1 strain, the antisera against recombinant proteins were produced by using guinea pigs. The result showed that the antisera neutralized the GC-1 strain. However, the neutralizing titer was higher in antisera against the VP2 gene expressed recombinant protein than that of VP2 partial gene recombinant protein.