• Title/Summary/Keyword: cELISA

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Effect of H-Y Antibody on in vitro Development of Mouse Embryos (H-Y항체의 처리가 생쥐수정란의 발달에 미치는 영향)

  • 고정재;심호섭;김종배;박홍양;정길생;이경광
    • Korean Journal of Animal Reproduction
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    • v.10 no.1
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    • pp.42-48
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    • 1986
  • These experiments were carried out to develop new techniques identifying XX-bearing embryos prior to implantation by immunological method. Antiserum to histocompatibility-Y(H-Y) antigen was prepared in adult SD(sprague-dawley) female rat by repeated immunization of newbone testis supernatant from males of the same strain. ELISA test was used to identify the H-Y antibody of antiserum. Total 124 mouse embryos (8-cell stage) were treated with H-Y antiserum and complement in BSA free Ho, pp. and Pitt's medium and cultured under the gas phase of 5% CO2 in air at 37$^{\circ}C$ for 24 to 48 hrs. The morphological characteristics of embryos treated were observed under the phase-contrast micro scope. The results obtained in these experiments were summarized as follows: 1. Optimal Density of H-Y antibody were a, pp.ared to be 0.27-0.47 by ELISA test. 2. Of total 124 embryos treated with H-Y antiserum and complement 69(55.6%) embryos developed to blastocyst and 55(44.4%) destroyed or arrested.

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Iridovirus infection of cultured juvenile flounder (Paralichthys olivaceus) in nursery (종묘장 넙치 치어에서의 Iridovirus 감염)

  • Kim, Tae-Jung;Jang, Eun-Jin;Kim, Jong-Su;Lee, Jae-Il
    • Korean Journal of Veterinary Research
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    • v.46 no.1
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    • pp.21-25
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    • 2006
  • Iridovirus is an icosahedral cytoplasmic double-stranded DNA virus with a genome size of 170-200kb. Outbreaks of fish iridovirus infection are characterized by their wide geographic distribution and broad host spectrum, especially in water temperatures of $25-27^{\circ}C$ Recently, the causative agent of high mortalities in flounder (Paralichthys olivaceus) was identified as fish iridovirus in Korea. Iridoviral infection repeatedly occurs in the same area for long periods, suggesting the possibility of viral infection in nursery. To examine this, the existence of iridovirus in juvenile flounders was detected by PCR using virus-specific primers. Antibodies induced against this virus were also examined using ELISA. As a result, juvenile fish in nursery were found to be previously infected with iridovirus, suggesting that proper prevention systems are required.

In Vitro Studies on the Release of Intracelluar Prolactin from Lymphocytes Using Strees Related Amines and Hormones

  • Sharma, G.T.;Majumdar, A.C.;Gupta, L.K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.12 no.7
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    • pp.1031-1034
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    • 1999
  • Circulating lymphocytes collected from control and heat-stressed buffaloes were subjected to in vitro culture with glucocorticoids, epinephrine or serotonin and their effect, if any, on the release of intracellular prolactin (PRL) was studied using ELISA and C-ELISA techniques. It was noted from the study that PRL level was higher in lymphocytes than in plasma of the control and heat-stressed animals, and that the PRL levels increased in the plasma of heat-stressed animals compared to that of non stressed animals with a significant decrease in lymphocytic PRL content by heat stress. Epinephrine and serotonin significantly increased the release of intracellular PRL from the lymphocytes of both in the control and the heat-stressed buffaloes but release of PRL from lymphocyte was not significantly changed by cortisol treatment in both control and heat-stressed buffaloes as compared to epinephrine and serotonin in vitro. When lympocytes were incubated with serotonin, it caused drastic lysis of the lymphocytes but epinephirine and cortisol did not show any lysis. It may be concluded from this study that hormones like epinephrine or serotonin known to increase during stress, release intracellular PRL from lymphocytes, the satellite PRL storage/synthesizing organ of blood, although the mechanism of the release is different.

Neutralizing Chimeric Mouse-human Antibodies against Burkholderia pseudomallei Protease: Expression, Purification and Characterization

  • Chan, Shzu-Wei;Ong, Guan-Im;Nathan, Sheila
    • BMB Reports
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    • v.37 no.5
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    • pp.556-564
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    • 2004
  • A recombinant Fab monoclonal antibody (Fab) C37, previously obtained by phage display and biopanning of a random antibody fragment library against Burkholderia pseudomallei protease, was expressed in different strains of Escherichia coli. E. coli strain HB2151 was deemed a more suitable host for Fab expression than other E. coli strains when grown in media supplemented with 0.2% glycerol. The expressed Fab fragment was purified by affinity chromatography on a Protein G-Sepharose column, and the specificity of the recombinant Fab C37 towards B. pseudomallei protease was proven by Western blotting, enzyme-linked immunosorbent assay (ELISA) and by proteolytic activity neutralization. In addition, polyclonal antibodies against B. pseudomallei protease were produced in rabbits immunized with the protease. These were isolated from high titer serum by affinity chromatography on recombinant-Protein A-Sepharose. Purified polyclonal antibody specificity towards B. pseudomallei protease was proven by Western blotting and ELISA.

Anti-inflammatory and Wrinkle Improvement Effects of Peptides from Ginseng Berry Amino Acidic Complex (진생베리 아미노산 복합체로부터 분리한 펩타이드의 항염, 주름개선 효과)

  • Kang, Sang Moon;Park, Chung
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.45 no.3
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    • pp.299-306
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    • 2019
  • Ginseng berry (GB) contains Ginsenoside Re and has anti-inflammatory and anti-wrinkle properties. In this study, TLC fractions 1, 2, and 4 of the ginseng berry amino acid complex were identified and analyzed by HPLC. And identified a peptide (AP-1) by LC/MASS analysis of fraction 1. The anti-inflammatory activity was confirmed by investigating the inhibitory effect of AP-1 on NO production. In addition, collagen synthesis using procollagen type I C-peptide (PIP) ELISA kit was 50% higher effective than that of the control group. From these results, the peptide isolated from ginseng berry amino acid complex is considered to have anti-inflammatory and anti-wrinkle effect, and may be useful as an anti-inflammatory and anti-aging cosmetic raw material.

Micrografting and Heat Treatment Combination for Eliminating Virus of CTV-infected Citrus (CTV 바이러스 보균 감귤나무로부터 열처리와 경정접목을 통한 바이러스 제거)

  • Chae, Chi Won;Yun, Su Hyun;Park, Jae Ho;Hyun, Jae Wook;Koh, Sang Wook;Lee, Dong Hoon
    • Journal of Life Science
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    • v.23 no.2
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    • pp.267-272
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    • 2013
  • This study was conducted to eliminate viruses from citrus-infected plants using micrografting and thermotherapy. Six citrus cultivars including a 'Setoka' hybrid were used as plant sources. The TAS-ELISA technique demonstrated that several plants were CTV positive. However, no CTV symptoms were detected in plants obtained from shoots and treated at a high temperature of $40^{\circ}C$ during the day and night and micrografted for two weeks with old trifoliate orange rootstock in vitro. Indexing of CTV, SDV, and CTLV for RT-PCR analysis of the eleven citrus seedlings, including 'Setoka', 'Samdajosang', 'Pungkwang', 'Shiranuhi', and 'Ehimekashi dai28go' was virus free following the micrografting and thermal therapy.

Suppressive effects of Th2 cytokines expression and the signal transduction mechanism in MC/9 mast cells by flavonol derived from Ginkgo biloba leaves (비만세포에서 은행잎 플라보놀에 의한 Th2 Cytokine 발현 및 신호전달 억제 기전 효과)

  • Kwon, Hae-Young;Chung, Kyu-Jin;Cheong, Kwang-Jo
    • Journal of Digital Convergence
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    • v.13 no.8
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    • pp.503-514
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    • 2015
  • The effects of Flavonol contents from Ginkgo biloba leaf on anti-atopy activity have not rarely been verified. This study is to investigate the effects of flavonol on Th2 cytokine production in MC/9 mast cells. For this, flavonol was analyzed by ELISA and Real-time PCR. Analysis results showed that flavonol significantly suppressed production of Th2 cytokines(IL-13, MIP-1a) in a dose dependent manner. The mRNA expression of IL-4, IL-5, IL-13, TNF-a were effectively restrained by Flavonol at the concentration 25,50,$100{\mu}g/m{\ell}$. And decrease of expression of NFAT-1, c-jun protein was confirmed by western blot analysis. These results indicate that flavonol has effects of decreasing the Th2 cytokine production in the MC/9 mast cell causing inhibition of transcription factors such as NFAT-1, c-jun. Thus, we would like to brief that flavonol may have the applicability as therapeutic agent for atopic dermatitis.

Detection of Antibodies Against SARS-Coronavirus Using Recombinant Truncated Nucleocapsid Proteins by ELISA

  • Lee, Hyun-Kyoung;Lee, Byoung-Hee;Dutta, Noton Kumar;Seok, Seung-Hyeok;Baek, Min-Won;Lee, Hui-Young;Kim, Dong-Jae;Na, Yi-Rang;Noh, Kyoung-Jin;Park, Sung-Hoon;Kariwa, Hiroaki;Nakauche, Mina;Mai, Le Quynh;Heo, Suk-Jin;Park, Jae-Hak
    • Journal of Microbiology and Biotechnology
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    • v.18 no.10
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    • pp.1717-1721
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    • 2008
  • Severe acute respiratory syndrome (SARS) is a life-threatening emerging respiratory disease caused by the coronavirus, SARS-CoV. The nucleocapsid (N) protein of SARS-CoV is highly antigenic and may be a suitable candidate for diagnostic applications. We constructed truncated recombinant N proteins (N1 [1-422 aa], N2 [1-109 aa], and N3 [110-422 aa]) and determined their antigenicity by Western blotting using convalescent SARS serum. The recombinants containing N1 and N3 reacted with convalescent SARS serum in Western blotting. However, the recombinant with N2 did not. In ELISA using N1 or N3 as the antigens, positive results were observed in 10 of to (100%) SARS-CoV-positive human sera. None of 50 healthy sera gave positive results in either assay. These data indicate that the ELISA using N1 or N3 has high sensitivity and specificity. These results suggest that the middle or C-terminal region of the SARS N protein is important for eliciting antibodies against SARS-CoV during the immune response, and ELISA reactions using N1 or N3 may be a valuable tool for SARS diagnosis.

GENE EXPRESSION FOR LYMPHANGIOGENIC FACTORS IN ORAL MUCOSAL SQUAMOUS CELL CARCINOMA (구강점막 편평상피세포암에서 림프관형성 유전자 발현)

  • Park, Young-Wook;Kim, Seong-Gon;Kim, So-Hee;Kim, Han-Seok;Kim, Min-Keun
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.31 no.6
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    • pp.453-460
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    • 2009
  • Background and Purpose: Vascular endothelial growth factor (VEGF)-C, VEGF-D and their tyrosine kinase receptor, VEGF receptor (VEGFR)-3 are recently known to have lymphangiogenic activities in various tumor types. Oral mucosal squamous cell carcinoma (OMSCC) easily metastasizes to cervical lymph nodes, so we determined the expression levels of VEGF-C, VEGF-D and VEGFR-3 in oral squamous cell carcinoma. Materials and Methods: We performed Western blot analyses with 4 OMSCC cultured tumor cell lines (SCC9, KB, YD-10B, YD-38), and with 7 surgical specimens of OMSCC for the detection of VEGF-C, VEGF-D and VEGFR-3 proteins. Expression of VEGF-C mRNA as well as mRNA for VEGFR-3 in 4 OMSCC cell lines (KB, SCC-4, SCC-9, YD-10B) was investigated by RT-PCR. We also measured VEGFC/VEGF-D protein concentrations in the media and protein concentration of VEGFR-3 in cell lysates of 4 OMSCC cell lines (SCC9, KB, YD-10B, YD-38) using commerical ELISA kits. Finally, we performed immunoprecipitation for the detection of VEGF-C in cell lysates of 4 OMSCC cells (KB, SCC-4, SCC-9, YD-10B) and real-time RT-PCR for the quantification of VEGF-C mRNA. Results: In the result of Western blotting with cell lysates of 4 OMSCC cells, we could not detect the protein expression of VEGF-C, VEGF-D, and VEGFR-3. But, all tumor tissues demonstrated VEGF-C and VEGFR-3. VEGF-C mRNA was detected at various levels in 4 OMSCC cell lines. Moreover, OMSCC cells secreted VEGF-C, not VEGF-D and VEGFR-3 was also detected in cell lysates of OMSCC by ELISA. Immunoprecipitation and real-time RT-PCR revealed VEGF-C was also expressed in 4 OMSCC cell lines. Conclusion: Taken together, tumor cells of OMSCC secrete VEGF-C, not VEGF-D. And VEGFR-3 is expressed tumor cells as well as OMSCC tumor tissues, needs further study.

Effect of M11C (Non-lectin Components) Obtained from Korean Mistletoe on the $IL-1\beta$ Secretion from Mouse Splenocytes (쥐의 비장세포로부터 $IL-1\beta$ 분비에 있어서 한국산 겨우살이 추출물 M11C (비렉틴 구성물질)의 효과)

  • Jun, Myung-Ha;Kang, Tae-Bong;Chang, Sung-Ho;Choi, Wahn-Soo;Seong, Nak-Sul;Her, Erk
    • Korean Journal of Medicinal Crop Science
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    • v.15 no.1
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    • pp.38-45
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    • 2007
  • Korean mistletoe (Viscum album L) extract has been found to posses immunoregulating activity. In this study, Korean mistletoe extract, M11C (non-lectin components), was used to know whether this extract activates splenocytes to secret interleukin $1\beta(IL-1\beta)$. The splenocytes were treated with M11C, and then collected the supernatant and cell lysate that were prepared to analyze the level of $IL-1\beta$, using ELISA, immunoblotting, and RT-PCR. Maximum effective dose and time of M11C on $IL-1\beta$ secretion from splenocytes were $200{\mu}g/m\ell$ and 8 hours, respectively. Treatment dose and time for the maximum expression of $IL-1\beta$ mRNA were $200{\mu}g/m\ell$ and 4 hours, respectively. Saccharide degradation enzyme Viscozyme L completely blocked the effect of M11C on $IL-1\beta$ secretion from splenocytes. As the result, among non-lectin components saccharide could be regarded as a main component for $IL-1\beta$ expression from splenocytes.