• 제목/요약/키워드: Serum Protein electrophoresis

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Proteomic Analysis of Serum Samples from Natural Herb and Immunoglobulin Yolksac (Ig Y) treated Porcine

  • Park, Hyeon-Soo;Nagappan, Arulkumar;Hong, Gyeong-Eun;Yumnam, Silvia;Lee, Ho-Jung;Kim, Gon-Sup
    • 한국임상보건과학회지
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    • 제1권1호
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    • pp.83-89
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    • 2013
  • Purpose. Natural herb has been used for traditional medicine for thousands of years. For this reason, alternative medicine methods affect to domestic animal field. The main purpose of this study was to confirm the regulated proteins after feed additive treatment. Methods. We used total thirty male pigs were used for this experiment. E. coli and Salmonella typhimurium, were used for promote enteritis. Animals were divided into a negative control group, positive control group and test group to determine the effect of an additive mixture on the changes in protein expression in serum. The pigs were given a food supplemented with a natural herbal additive containing immunoglobulin yolksac (IgY) at concentrations 1%. On the 1st week and after eight weeks of feeding, the serums were collected from each group. The changes in the serum proteome as a response to the herbal additive were examined using two-dimensional polyacrylamide gel electrophoresis. Results. A total of 14 differentially expressed protein spots were identified by comparing the protein profiles of the control and additive treated porcine serums. Finally, 7 proteins were detected by MALDI-TOF/MS. Moreover, the proteins detected are involved in a cholesterol control factor, inflammation regulator, Conclusion. These results support of the hypothesis that a natural herbal additive containing IgY can affect the immune regulation system and reduce the pathological process by microbial infections.

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非姙娠 및 姙娠한 女子의 血淸蛋白質 패턴의 比較 (The Comparison of Protein Patterns of Sera in Non-Pregnant and Pregnant Women)

  • Ha, Man-Joon;Park, Won-Chul
    • 한국동물학회지
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    • 제29권2호
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    • pp.86-106
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    • 1986
  • 사람의 혈액중 혈청으로써 임신의 진행에 따른 단백질의 변화 양상을 조사함에 있어서 정상인 남자, 비임신 여자, 임신한 여자 및 분만 직후 여자의 혈청을 SDS/polyacrylamide gel 전기영동과 2차원 전기영동 및 아미노산분석을 시도하였다. 각 실험 방법에 의해 분석된 결과는 다음과 같다. 1. SDS/polyacrylamide gel 전기영동에 의해 분자량 약 10,000에서 110,000 dalton 사이의 정상인 남자와 비임신 여자를 비교하였을 때의 숫적 양상은 서로 동일하였으나 bands 3 (22,000 dalton)과 6(39,000)등은 남자에 있어서 여자보다 적은 양으로 나타났다. 비임신 여자와 2주 간격으로 혈액을 채취한 임신한 여자의 혈청단백질 패턴을 비교하면 여러 band들의 양적인 증감이 관찰되었다. 임심한 여자의 단백질은 비임신에 비해 band 3(22,000)이 16주까지는 비임신의 경우와 거의 비슷한 양상을 보였으나 18주부터 임신 말기까지는 단백질의 양이 계속하여 감소되었다. 반면 bands 4(24,000), 9(69,000), 10(70,000), 12(80,000), 13(83,000), 14(86,000), 15(91,000) 및 16(94,000)은 임신이 진행됨에 따라 일반적으로 증가되나 그 이후에는 약간씩 감소하였다. 분만 직후의 혈청단백질의 패턴을 임신 말기와 비교하였을 때 bands 12(80,000), 15(91,000) 및 16(94,000)들의 양이 비교적 증가하였고 여아를 분만한 경우 남아를 분만한 경우보다 bands 4(24,000), 7(51,000) 및 10(70,000)들의 단백질 양이 많음이 관찰되었다. 2. 2차원 전기영동으로 남자와 비임신 여자의 혈청단백질 패턴을 비교하면 spot a(22,000)의 3개의 spot가 남자에 있어서는 나타나지 않았고, spotc(39,000)의 군은 남자에 있어 여자보다 농도가 매우 낮았다. 임신한 여자에 있어 albumin이 임신 10주와 12주에 매우 감소하였고 그 이후에 다시 회복되었다. 그리고 spot f(70,000)는 임신 10주에 매우 감소 되었다가 그 이후에 증가되었다. 3. 각 군들의 아미노산을 분석한 결과 임신한 여자에 있어서 전반적으로 glutamic acid, lysine, aspartic acid, leucine 및 valine이 대체로 많은 양으로 나타났고, methionine, isoleucine 및 glycine이 대체로 적은 양을 가지는 것으로 나타났다. 전체 아미노산의 양은 임신중기에 현저히 증가하였다가 말기에는 중기에 비해 감소되었다. 이상의 결과를 볼 때 임신기간 중에는 모체의 혈청을 구성하는 단백질이 특정한기에 따라 증가하므로 임신한 여자의 혈청단백질을 조사함으로써 임신의 각기에 나타나는 독특한 단백질 양상 변화를 밝혀내는 동시에, 앞으로 좀더 연구함으로써 태아의 성을 단백질 패턴으로 명확히 구별해 낼 수 있을 것으로 생각된다.

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Gradient 2-D PAGE를 이용한 양수 프로테옴 분석 (Proteome Analysis of Amniotic Fluid by gradient 2-D PAGI)

  • 이은희;김재찬;변상요
    • KSBB Journal
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    • 제18권1호
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    • pp.35-38
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    • 2003
  • 양수 내에 존재하는 총 단백질을 이차원 전기영동을 이용하여 분리 분석하였고, gradient gel을 이용하여 양수 내에 소량으로 존재하는 미세 단백질까지 분리하였다. 양수 내에는 고농도로 존재하는 단백질이 있는데 이것이 serum albumin precursor임을 확인하였고, 8-18% gradient gel의 이용으로 분해능(resolution)이 향상되어 미세 단백질을 분리 분석할 수 있었다. 이차원 전기영동 후 MALDI-TOF를 이용하여 단백질을 identification하여 기존의 양수 protein database에 존재하는 단백질을 확인하였고, 존재하지 않는 새로운 단백질을 분리 분석하였다.

Significance of Oligoclonal Bands after Stem Cell Transplantation in Multiple Myeloma Cases

  • Liu, Ai-Jun;Zong, Hong;Yang, Guang-Zhong;Zhai, Yu-Hua;Li, Li-Hong
    • Asian Pacific Journal of Cancer Prevention
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    • 제13권4호
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    • pp.1483-1486
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    • 2012
  • Objective: To determine the characteristics of oligoclonal bands that are frequently detected by serum protein electrophoresis (SPEP) and immunofixation electrophoresis (IFE) after stem cell transplantation. Methods: We retrospectively analyzed 56 patients with multiple myeloma (MM) undergoing transplantation, and standard immunofixation electrophoresis was used to identify and quantify paraproteins. Results: The median follow-up was 35 months (range, 10-76months) and 21 patients relapsed. Twelve (25.0%) demonstrated oligoclonal bands after a median time 1.4 months (range, 1-3months), with a median duration of 5.8 months (range, 1-15months). The majority patients with oligoclonal bands had normal quantities of immunoglobulins and the one year event free survival (EFS) was 92%, even higher than for patients without OBs (P=0.002). Conclusion: Oligoclonal bands frequent develop post-transplantation in MM cases. In the vast majority of patients, they may not represent relapsed disease, and more likely represent a transient phenomenon representing recovery of impaired immunoglobulin production.

한국재래산양 혈청단백질 좌위의 유전적 다형현상에 관한 연구 (Genetic Polymorphisms of the Serum Protein Locus in Korean Native Goats)

  • 김계웅
    • 한국가축번식학회지
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    • 제19권3호
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    • pp.235-241
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    • 1995
  • These experiments were carried out to find genetic polymorphisms of Serum protein like Pre-albumin(Pr), Albumin(Al) and Transferrin(Tf), and establish preservation of pure pedigree in Korean Native Goats(KNG). Their serum was collected and examined from the total of 74 KNG that raised in Tang Jin district, Chungnam-province. They were biochemically analysed by polyacrylamide gel(7.5%) electrophoresis(PAGE) in order to estimate the frequencies of genotypes and alleles existing on each trait locus. The results obtained in these experiments were summarized as follows ; 1. In the serum Pre-albumin(Pr) locus, the frequencies of genotypes for hetero AB and homo BB observed were 55.4%, and 44.6%, respectively. While homo AA was not found in the Pr locus. The frequencies of gene in PrA and PrB were 0.723 and 0.277, respectively. Accordingly, the Pr loci were assumed to be controlled by alleles PrA and PrB. 2. The frequencies of genotypes of homo BB and hetero AB detected in Albumin(Al) locus were 75.7% and 24.3%, respectively. However, AA type was not observed in the Al locus. The gene frequencies of AlA an AlB were 0.879 and 0.121, respectively. Also, the Al loci were considered to be controlled by alleles AlA and AlB. 3. The frequencies of genotypes for hetero AD and homo DD found in Transferrin (Al) locus were 79.7% and 20.3%, respectively. Whereas, homotype AA was not detected in this locus. The gene frequencies of TfA and TfD were 0.399 and 0.601, respectively. Therefore, the serum Tf loci were assumed to be controlled by alleles Tfa and Tfd.

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Detection of Retinol-binding Protein in Bovine Yolk Sac, Chorion and Allantois by Immunoperoxidase Method

  • Liu, Kaung Huei
    • Asian-Australasian Journal of Animal Sciences
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    • 제15권6호
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    • pp.783-788
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    • 2002
  • Bovine yolk sac at day 24 of pregnancy, and placental membranes (chorion and allantois) from days 70 and 100 of pregnancy were isolated and cultured in a modified minimum essential medium in the presence of $[^{35}S]$methionine. Proteins synthesized and secreted by isolated bovine yolk sac, chorion and allantois were analyzed by fluorography of two-dimensional polyacrylamide gel electrophoresis. Serum-like proteins,transferrin, ${\alpha}$-fetoprotein, ${\alpha}$1-antitrypsin and ${\alpha}$1-acid glycoprotein,were the major protein products of yolk sac. A 21 kDa protein produced by yolk sac was identified immunochemically as retinol-binding protein (RBP). Chorion and allantios from days 70 and 100 of pregnancy were active in protein synthesis and secretion. Both chorion and allantois did not secret serum-like proteins but secreted a number of neutral-to-acidic proteins including RBP. Secretory proteins produced by the yolk sac, chorion and allantois may play important roles in the embryonic development and the successful outcome of pregnancy. Antiserum against bovine placental RBP was employed to the immunocytochemistry by immunoperoxidase method. Immunoreactive RBP was localized in epithelial cells and island-like cell clones of yolk sac. Immunostaining for RBP was detected in simple columnar epithelium of chorion and in simple squamous epithelium of allantois. In the present study, proteins synthesized and secreted by yolk sac at day 24 of pregnancy, chorion and allantois from days 70 and 100 of pregnancy were characterized In addition, RBP was localized in yolk sac, chorion and allantois by immunoperoxidase method. The immunoperoxidase method has been proven to be a very effective technique to identify the cellular source of protein synthesis in extraembryonic membranes.

Protective Ability of Ethanol Extracts of Hypericum scabroides Robson & Poulter and Hypericum triquetrifolium Turra against Protein Oxidation and DNA Damage

  • Kizil, Goksel;Kizil, Murat;Ceken, Bircan
    • Food Science and Biotechnology
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    • 제18권1호
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    • pp.130-136
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    • 2009
  • The present study was conducted to determine the protective ability of the ethanol extracts of Hypericum scabroides Robson & Poulter (HS) and Hypericum triquetrifolium Turra (HT) against the protein oxidation and DNA damage induced by Fenton system. The ability of HS and HT to prevent oxidative damage to bovine serum albumin (BSA) induced by $Fe^{3+}/H_2O_2$ and ascorbic acid was investigated. The ethanol extracts of HS and HT at different concentrations ($50-1,000{\mu}g/mL$) efficiently prevented protein oxidation induced by hydroxy radical as assayed by protein oxidation markers including protein carbonyl formation (PCO) and polyacrylamide gel electrophoresis. The effect of ethanol extracts of HS and HT on DNA cleavage induced by UV-photholysis of $H_2O_2$ using pBluescript M13+ plasmid DNA were investigated. These extracts significantly inhibited DNA damage induced by reactive oxygen species (ROS). Therefore, HS and HT extracts may be useful in the food industry as effective synthetic antioxidants.

Characterization of gender-specific bovine serum

  • Kim, Ji-Hoe;Kim, Min-Soo;Nahm, Sang-Soep;Lee, Dong-Mok;Pokharel, Smritee;Choi, In-Ho
    • Animal cells and systems
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    • 제15권2호
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    • pp.147-154
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    • 2011
  • Animal cell cultures generally require a nutrient-rich medium supplemented with animal serum. Adult bovine serum contains a variety of nutrients including inorganic minerals, vitamins, salts, proteins and lipids as well as growth factors that promote animal cell growth. To evaluate the potential use of gender-specific bovine serum (GSBS) for cell culture, the biochemical properties of male serum (MS), female serum (FS) and castrated-male serum (CMS) were investigated. Overall, the chemical profile of GSBS was similar to that of bovine references except for glucose, creatine kinase, lactate dehydrogenase and potassium. FS showed elevated total protein and sodium concentrations compared to MS and CMS. Proteins present in MS, FS and CMS but absent in fetal bovine serum (FBS) were selected by two-dimensional gel electrophoresis and identified by peptide mass fingerprinting. Some of the identified proteins are known to be involved in immune responses and the others have unknown physiological roles. Moreover, it was found that some proteins such as alpha-2-macroglobulin appeared to be gender-specific with higher contents in FS. Insulin and testosterone was significantly higher in MS, and $17{\beta}$-estradiol and estrone were higher in FS, as compared to the other sera. Taken together, the results indicate that each GSBS has a different ratio of components. Differences in serum constituents may affect cell cultures in a different manner and could be beneficial, depending on the specific aim of cell cultures.

이차원적 전기영동을 이용한 홍화씨의 신생골 형성 기작에 관한 연구 (Possible Process of Safflower Seed on New Bone formation by 2-Dimensional Electrophoresis)

  • 라도경;정태성;김종수;송해룡;김용환;강정부;강호조;연성찬;신기욱
    • 한국임상수의학회지
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    • 제19권1호
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    • pp.49-54
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    • 2002
  • Korean safflower (Carthami Flos) seed has been known to have healing effects on both bone fracture and osteoporosis. On the base of such a notice, this experiment was carried out to explore the effects of safflower seed on bone formation and bone repair. In addition, the healing mechanism was evaluated by analysing serum after feeding the seed to experimental. animals. The effect of Korean safflower seed were evaluated with 40 rats,3-month old. Forty Sprague-Dawley rats composed of 20 male and 20 female were underwent unilateral tibial defect and then fastened with unilateral fixators. The operated rats were divided into two groups depending on the composition of diet, such as positive control group fed normal diet (C-OP group) and safflower seed group fed 30% of safflower seed diet and 70% of normal diet (S-OP group). Postoperative radiographys were taken once in 2 weeks to evaluate callus formation for operated groups. In addition, a possible protein spots involved in bone recovery were examined using 2-Dimensional Gel Electrophoresis (2-DE). The comparison of the radiography between C-OP and S-OP group were showed that the safflower seed diet appeared to stimulate the formation of callus in the rat. On the images of 2-BE, it was able to identify possible five protein spots, having pl from 4 to 5 and molecular weight range from 24 to 26 kDa, involved in bone formation and repair, since no differing protein spots were found the two between groups except the five spots. No differences were observed between two groups before operation, but clear and bigger protein spots were observed from the S-OP group compared with C-OP group on 6 and 9 weeks post operation. These protein spots were, however, showed similar sizes and densities between two groups in 12 weeks later. The transformation of protein spots was suggested that these protein spots were involved in bone formation and recovery, in addition safflower seed might induce the formation of factors and activate these factors. In conclusion, this study suggest that safflower seed influence a variety of factors in the course of bone formation or the periods of remedy.