• Title/Summary/Keyword: Blotting

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A Study on the Measurement of Water Content in Hydrogel Contact Lens by Gravimetric Method (중량측정법에 의한 하이드로젤 콘택트렌즈의 함수율 측정에 관한 연구)

  • Kim, Tae-Hun;Ye, Ki-Hun;Sung, A-Young
    • Journal of Korean Ophthalmic Optics Society
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    • v.13 no.4
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    • pp.59-63
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    • 2008
  • Purpose: The purpose of this study is to understand a relation of water content measurement with two different method, water removal method and dry method. Methods: 72 hydrogel contact lens containing various water content (ranged from 47% to 58%) were measured by the gravimetric method at 20$^{\circ}C$and 21% of the humidity. We weighed the dried test specimen at room temperature for 30 min after cooling. Results: In dry blotting method, the water content was measured to 47.43${\pm}$8.48%. The water contents was measured to 48.15${\pm}$8.36% with wet blotting condition. It was found that wet blotting method showed the higher water content of about 7% than dry blotting method. In water content with two dry methods, each of results was measured by 47.89${\pm}$8.06% and 49.56${\pm}$7.06%. In case of microwave method, the water content was measured significantly higher water content of about 1.67% than vacuum oven method. However, no statistical difference was found (p>0.05). Conclusions: In water removal method (Dry blotting method and Wet blotting method) to weigh hydrated test specimen, wet blotting method showed significantly higher water content than dry blotting method. Also in case of dry methods (vacuum oven and microwave) to weigh dry test specimen, water content of microwave method showed higher water content than vacuum oven method, but it should be noted that microwave oven method must be used carefully to measure accurateness on the specimen position and wave power.

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Reliability of Measurements of Back Vertex Power for Soft Contact Lenses Using an Auto-Lensmeter (자동렌즈미터를 이용한 소프트 콘택트렌즈의 굴절력 측정 방법에 관한 신뢰도)

  • Kim, Kun-Kyu;Lee, Wook-Jin;Lee, Sun-Haeng;Kwak, Ho-Won;Yu, Dong-Sik
    • Journal of Korean Ophthalmic Optics Society
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    • v.15 no.1
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    • pp.39-46
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    • 2010
  • Purpose: To assess the reliability for measuring the back vertex power of soft contact lenses by dry blotting and wet cell method using an auto-lensmeter. Methods: The soft contact lenses used for measurement were 5 types that were distributed in Korea, and 4 back vertex powers (-1.50D, -3.00D, -6.00D, -9.00D) were used. and repeatability and reproducibility were evaluated by measuring them with an auto-lensmeter by two examiners. Results: Measured powers by dry blotting method were ranged in mean differences from 0.03D to 0.18D for overall lenses, 0.10D to 0.18D for silicone hydrogel lenses, 0.03D to 0.08D for hydrogel lenses. The mean differences between two examiners were less than 0.10D, and the inter-examiner reproducibility was good for dry blotting method. The mean difference between powers determined by wet cell method were 0.09D to 0.69D, the mean differences between two examiners were 0.02D to 0.59D. The reliability of measurements and inter-examiner reproducibility were less than dry blotting method. Conclusions: The reliability of measurements for all materials was better in dry blotting than wet cell method, the re liability of measurements for silicone hydrogel lenses was low in both methods. In clinical practical which requires quick checking of back vertex power using an auto-lensmeter. dry blotting method is thought to be more efficient than wet cell one.

Improvement of Antigen Blotting in a Tissue Blot Immunobinding Assay for the Detection of Two Chili Pepper Viruses

  • Han, Jung-Heon;Shin, Jun-Sung;Kim, Young-Ho;Kim, Byung-Dong
    • Journal of Microbiology and Biotechnology
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    • v.17 no.11
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    • pp.1885-1889
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    • 2007
  • The tissue blot immunobinding assay (TBIA) is widely used for the detection and localization of plant viruses in various plant tissues. The basic experimental procedures of TBIA sampling and blotting were simplified using commercially available micropipette tips. This method was termed the ring-blot immunobinding assay (R-BIA), as the blot on the membrane forms a ring shape. The detection efficacy of R-BIA was tested for two chili pepper viruses, pepper mild mottle tobamovirus (PMMoV) and pepper mottle potyvirus (PepMoV), following the optimized serological procedures of TBIA (length of the incubation period and BSA concentration, and primary and secondary antibodies). Sensitivity of the R-BIA was about 1 ng/ml of purified PMMoV in pepper leaf sap from a healthy pepper plant. R-BIA also showed high specificity in the detection of PMMoV and PepMoV. Moreover, the modified sampling and blotting procedures were simpler and more reliable than other TBIA methods (such as whole-leaf blotting and crushed-leaf blotting), suggesting that the R-BIA may be used for medium- to large-scale detection of plant viruses in laboratories with minimal facilities.

Ionizing Radiation Effect on the Carbohydrate Moiety of Chicken Ovomucoid (계란 ovomucoid의 탄수화물 부분에 미치는 이온화방사선의 영향)

  • Lee, Young-Keun;Kim, Jin-Kyu;Kim, Jae-Sung;Song, Hi-Sup;Charoen, Saovapong;Amornraksa, Kitti
    • Journal of Radiation Protection and Research
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    • v.22 no.1
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    • pp.23-27
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    • 1997
  • Radiation effects on carbohydrate moiety of chicken ovomucoid, a protease inhibitor as a typical allergenic glycoprotein of egg white, was observed. The trypsin inhibitory activity of chicken ovomucoid decreased exponentially and the inactivation was more significant irradiated in $N_2$ than in $O_2$. From the protein blotting, radiation caused protein degradation in $O_2$ and protein aggregation also in $N_2$. The patterns of carbohydrate blotting were also similar with that of protein blotting. Sugar chains in low molecular weight fraction (MW<5,000) were released by radiation and those in $O_2$ were higher than in $N_2$. From the HPLC patterns of the degradation of sugar chains, all peaks of oligosaccharides have the tendency to decrease with the increase of radiation dose and more remarkable in $O_2$ than in $N_2$. These results suggest that ionising radiation could cause the overall conformational changes of ovomucoid by the degradation and release of oligosaccharides.

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Expression and Antibacterial Activity of Recombinant Human Lactoferrin in Methylotrophic Yeast, Pichia pastoris (Methylotrophic Yeast, Pichia pastoris에서 사람 락토페린의 발현 및 항균성 연구)

  • Lee Sang O;Im Eun Mi;Nam Eun Joo;Lee Hyune Hwan
    • Korean Journal of Microbiology
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    • v.40 no.4
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    • pp.348-354
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    • 2004
  • The expression and antibacterial. activity of recombinant human lactoferrin (hLf) was studied from meth­ylotrophic yeast Pichia pastoris. The gene encoding hLf, isolated from human breast cDNA library, was subcloned into the expression vector, pPIC3.5K under the control of AOX1 promoter. The gene was integrated into the host chromosome and was identified by Southern blotting. The expression of the integrated gene was investigated by RT-PCR, Northern blotting, SDS-PAGE and Western blotting. Discrete band corresponding to hLf was detected from the SDS-PAGE, which was confirmed by Western blotting. The expression was also confirmed by RT-PCR and Northern blotting. The antibacterial activity of the recombinant hLf (rhLf) was investigated using Staphy­lococcus aureus ATCC 6538P and Micrococcus flavus ATCC 10240 as test organisms. The rhLf showed strong antibacterial activities against the bacteria. Furthermore, many Gram-negative animal pathogens such as E.coli ATCC8739, 25922, and Salmonella typhimurium 114 and 115, Pseudomonas fluorescens ID 963 I, P. aeruginosa KCCM 11802, and Gram-positive bacteria Bacillus mesentericus were also inhibited in their growth by the rhLf.

Automated Protein-Expression Profiling System using Crude Protein Direct Blotting Method

  • Kobayashi, Hironori;Torikoshi, Yasuhiro;Kawasaki, Yuko;Ishihara, Hideki;Mizumoto, Hiroshi
    • 제어로봇시스템학회:학술대회논문집
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    • 2003.10a
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    • pp.2356-2361
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    • 2003
  • Proteome research in the medical field is expected to accelerate the understanding of disease mechanism, and to create new diagnostic concept. For protein profiling, this paper proposes a new methodology named CPDIB (Crude Protein Direct Blotting). In the CPDIB procedure, crude protein sample is directly immobilized on a membrane and the expression of protein molecules in the sample are analyzed quantitatively by using a special device called ImmobiChip, where the membrane is used as a field of the immune reaction. The over-all structure of the ImmobiChip is based on the conventional Slot blot device. Mechanical improvement in the air-tightness of the case holding the membrane realizes the direct blotting and results in high performance of stability in the immune reaction. In the measurement of multiple proteins, a dispensing robot is used for increasing the efficiency of handling of liquid. Cooperation of the dispensing robot with the ImmobiChip for immobilizing proteins realizes automated and stable performance of the CPDIB procedure. This paper shows the evaluation of the air-tightness of the ImmobiChip, the ability of analyzing proteins using the CPDIB procedure and the performance of the automated equipment.

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Utilization of pollen grains for the expression of epidermal growth factor (Epidermal growth factor 발현을 위한 화분립의 이용)

  • Choi, Byung-Jin;Park, Hee-Sung
    • KSBB Journal
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    • v.23 no.5
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    • pp.460-462
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    • 2008
  • Pollens grains collected from fully dehisced lily (Lilium longiflorum) anthers were given wounds by means of shaking in the presence of aluminum oxide particles. They were transformed by infiltration with Agrobacterium cells harboring a synthetic DNA encoding signal peptide-fused epidermal growth factor (EGF). After incubation for 24 hr in vitro, the pollen culture showed that EGF mRNAs and proteins were successfully expressed in the analysis of cDNA blot hybridization and immuno-blotting.

Overproduction and Operator DNA-Protein Blotting of R100 Mutant MerR from Shigella flexneri

  • Yoon, Kyung-Pyo
    • Journal of Microbiology and Biotechnology
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    • v.4 no.4
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    • pp.250-255
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    • 1994
  • Wild-type and four mutant R100 merR genes were cloned and the proteins overproduced under tac promoter control of pKK223-3. His118Ala, Cys117Ser, Cys126Ser, and wild-type MerR were successfully overproduced although amino-terminal 14 amino acids deletion mutant MerR was not successful. The amount of overproduced wild-type MerR protein as well as other mutant MerR was between 15%-20% of the total protein. The protein was able to be purified up to 95% homogeneity. Specific DNA-protein blotting experiments showed that the 95 bp operator containing DNA fragment could bind to Cys126Ser, His118Ala, and wild- type MerR, but not to Cys117Ser. These results were consistent with the previously reported complementation experiment results that His118Ala, Cys126Ser, and wild-type MerR could repress the mer operon but Cys117Ser could not.

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Expression of Extracellular Superoxide Dismutase Protein in Diabetes

  • Kim, Chul Han
    • Archives of Plastic Surgery
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    • v.40 no.5
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    • pp.517-521
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    • 2013
  • Background Diabetes is characterized by chronic hyperglycemia, which can increase reactive oxygen species (ROS) production by the mitochondrial electron transport chain. The formation of ROS induces oxidative stress and activates oxidative damage-inducing genes in cells. No research has been published on oxidative damage-related extracellular superoxide dismutase (EC-SOD) protein levels in human diabetic skin. We investigated the expression of EC-SOD in diabetic skin compared with normal skin tissue in vivo. Methods The expression of EC-SOD protein was evaluated by western blotting in 6 diabetic skin tissue samples and 6 normal skin samples. Immunohistochemical staining was also carried out to confirm the EC-SOD expression level in the 6 diabetic skin tissue samples. Results The western blotting showed significantly lower EC-SOD protein expression in the diabetic skin tissue than in the normal tissue. Immunohistochemical examination of EC-SOD protein expression supported the western blotting analysis. Conclusions Diabetic skin tissues express a relatively small amount of EC-SOD protein and may not be protected against oxidative stress. We believe that EC-SOD is related to the altered metabolic state in diabetic skin, which elevates ROS production.

Identification of Chicken Pork and Beef Meats by Chicken Specific Antibody (계육 특이항체를 이용한 원료육 단백질의 검색)

  • 임태진
    • Food Science of Animal Resources
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    • v.18 no.2
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    • pp.149-156
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    • 1998
  • Chicken beef pork meats and isolated soy protein (ISP) were heated at 10$0^{\circ}C$ for 30min and then heat-resistant proteins were fractionated to examine cross-resistant protein from chicken meat but not with beef pork or ISP. Dot blotting using the polyclonal antibody showed that the sen-sitivity for detecting chicken meat was 1$\mu$m and antibody-antigen reaction was dose-dependant. Results of dot blotting analysis to compare the amount of chicken meat present in arket meat products(Kentucky Frank sausage;chicken meat 46.52% and pork 24.92% vs Bulgogi Ham;chicken meat 28.89% and turkey 31.44%)showed that the significant differences between two meat products in terms of chicken meat concentrations. Dose-dependant dot-blotting reaction was also observed in chicken meat samples with various dilution.

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