• Title/Summary/Keyword: Neutral Red

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Fast Protein Staining in Sodium Dodecyl Sulfate Polyacrylamide Gel using Counter ion-Dyes, Coomassie Brilliant Blue R-250 and Neutral Red

  • Choi, Jung-Kap;Yoo, Gyurng-Soo
    • Archives of Pharmacal Research
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    • v.25 no.5
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    • pp.704-708
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    • 2002
  • A fast and sensitive protein staining method in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) using both an acidic dye, Coomassie Brilliant Blue R-250 (CBBR) and a basic dye, Neutral Red (NR) is described. It is based on a counter ion-dye staining technique that employs oppositely charged two dyes to form an ion-pair complex. The selective binding of the free dye molecules to proteins in an acidic solution enhances the staining effect of CBBR on protein bands, and also reduces gel background. It is a rapid staining procedure, involving fixing and staining steps with short destaining that are completed in about 1 h. As the result, it showed two to fourfold increase in sensitivity comparing with CBBR staining. The stained protein bands can be visualized at the same time of staining.

Change of Neutral Ginsenoside Contents in Red and Fresh Ginseng (Panax ginseng C. A. Meyer) by Hydrolysis (가수분해 처리에 의한 홍삼과 인삼의 중성 Ginsenoside 함량 변화)

  • Han, Jin Soo;Lee, Gang Seon;Tak, Hyun Seong;Kim, Jung-Sun;Ra, Jeong Woo;Choi, Jae Eul
    • Korean Journal of Medicinal Crop Science
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    • v.22 no.1
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    • pp.23-31
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    • 2014
  • This study was carried out to investigate change of ginsenoside contents in red and fresh ginseng according to root part and age by hydrolysis. Neutral total ginsenoside contents by hydrolysis in 6-year main root and lateral root were significantly increased than those by non-hydrolysis, as 41.6 and 32.8%, respectively. However, there was no significant difference in red ginseng. In fresh ginseng, ginsenoside contents of the protopanaxatriol group such as Re, Rf, $Rg_1$, $Rg_2$, and $Rh_1$ were not significantly different, but $Rb_1$, $Rb_2$, $Rb_3$, Rc, and Rd showed significant difference. The increase rate of neutral total ginsenoside content by hydrolysis was higher in epidermis-cortex than stele. Also, the neutral total ginsenoside content was fine root > rhizome > lateral root > main root, respectively. While there was no tendency towards the increase of ginsenoside by hydrolysis with the increase of root age in fine root and rhizome, there was significant decrease in main root and lateral root.

Electrochemical Control of Metabolic Flux of Weissella kimchii sk10: Neutral Red Immobilized in Cytoplasmic Membrane as Electron Channel

  • PARK, SUN-MI;KANG, HYE-SUN;PARK, DAE-WON;PARK, DOO-HYUN
    • Journal of Microbiology and Biotechnology
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    • v.15 no.1
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    • pp.80-85
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    • 2005
  • Electrochemical control of the metabolic flux of W. kimchii sk10 on glucose and pyruvate was studied. The growing cell of W. kimchii sk10 produced 87.4 mM lactate, 69.3 mM ethanol, and 4.9mM lactate from 83.1mM glucose under oxidation condition of the anode compartment, but 98.9 mM lactate, 84.3mM ethanol, and 0.2 mM acetate were produced from 90.8 mM glucose under reduction condition of the cathode compartment for 24 h, respectively. The resting cell of W. kimchii sk10 produced 15.9 mM lactate and 15.2 mM acetate from 32.1 mM pyruvate under oxidation condition of the anode compartment, and 71.3 mM lactate and 3.8 mM acetate from 79.8mM pyruvate under reduction condition of the cathode compartment. The redox balance (NADH/$NAD^+$) of metabolites electrochemically produced from pyruvate was 1.05 and 18.76 under oxidation and reduction conditions, respectively. On the basis of these results, we suggest that the neutral red (NR) immobilized in bacterial membrane can function as an electron channel for the electron transfer between electrode and cytoplasm without dissipation of membrane potential, and that the bacterial fermentation of W. kimchii sk10 can be shifted to oxidized or reduced pathways by the electrochemical oxidation or reduction, respectively.

Comparison of Intramuscular Fat Composition of Red Muscle and White Muscle (Red Muscle과 White Muscle의 근섬유간 지방질의 조성 비교)

  • Yang, Ryung;Kim, Kee-Tae;Shin, Wan-Chul
    • Korean Journal of Food Science and Technology
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    • v.21 no.4
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    • pp.505-510
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    • 1989
  • Red muscle and white muscle were separated from bovine, porcine and poultry skeletal muscles, respectively. Intramuscular lipids were extracted and fractionated to neutral-, glyco- and phospho-lipid by silica gel chromagraphy and then fatty acid composition were analyzed with gas chromatography. The results obtained were as follows; Total lipid content of red muscle was higher than that of white muscle in case of beef and chicken. In pork, however, total lipid content of white muscle was higher than red muscle The content ratio of neutral lipid to phospholipid revealed a number of distinctions between red and white muscle among animals. There were noteworthy differences in respect of polyunsaturated fatty acid. The intramuscular fat of pork had the higher content of highly polyunsaturated fatty acid such as arachidonic acid in contrast to beef.

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Comparison of Physicochemical Properties and Malonyl Ginsenoside Contents between White and Red Ginseng (백삼 및 홍삼의 이화학적 특성 및 말로닐 진세노사이드 함량 비교)

  • Oh, Myeong Hwan;Park, Young Sik;Lee, Hwan;Kim, Na Young;Jang, Young Boo;Park, Ji Hun;Kwak, Jun Young;Park, Young Soon;Park, Jong Dae;Pyo, Mi Kyung
    • Korean Journal of Pharmacognosy
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    • v.47 no.1
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    • pp.84-91
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    • 2016
  • Korean ginseng (Panax ginseng C. A. Meyer) has been used as a traditional herbal medicine in East Asia and is very popular in the world, because of its health benefits. To comparison of pharmacological components and physiochemical properties between white and red ginseng from same body, we analyzed ginsenoside and malonyl ginsenoside, ash, crude lipid/protein, fatty acid, mineral contents, total/reducing sugar, and total phenolic and acidic polysaccharide contents. The general components did not show any significant difference between white and red ginseng. Whereas, the content of neutral ginsenoside $Rb_1$, $Rb_2$, Rc and Rd were higher in red ginseng than those of white ginseng. However, malonyl ginsenoside such as $m-Rb_1$, $m-Rb_2$, m-Rc and m-Rd in white ginseng were similar to neutral ginsenoside $Rb_1$, $Rb_2$, Rc and Rd in white ginseng and far higher than those of red ginseng. These results exhibit that malonyl ginsenosides were converted to neutral ginsenosides in steaming process for red ginseng. So, we suggest that malonyl ginsenoside are necessary to applies in ginsenoside analysis of Korean ginseng.

A Study on the Cytotoxic Effect of Heavy metals (Cd, Ni, Zn) on Cultured Mouse Fibroblast L929 Cell line (생쥐 배양섬유 모세포주 L929에 미치는 중금속(Cd, Ni, Zn)류의 세포독성에 관한 연구)

  • 이종빈;나명석;황영진;위성욱;최진희;김선희;유춘만;김재민
    • Journal of Environmental Health Sciences
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    • v.23 no.2
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    • pp.98-105
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    • 1997
  • The study on the cytotoxicity of heavy metals was carried out to evaluate the cytotoxic effect of those on mouse L929 fibroblast cell in 96-well microtiter plates. The cytotoxicity was assayed by the neutral red, tetrazolium MTT, total protein, micronuclei test. The cytotoxicity of the heavy metals by neutral red and tetrazolium MTT was showed in order, cadmium > zinc > nickel for the cationic metals tested. The effect of metal-metal interaction on the cytotoxicity showed a marked reduction of cadmium toxicity by zinc, to a lesser degree, by nickel. The amount of total protein in treated group added heavy metals was less than that of the control and treated cadmium alone was less than those of combination with nickel or zinc. At midpoint cytotoxicity values of heavy metals, the frequency of micronuclei on the cell treated heavy metals was more than that of control and treated cadmium alone was more than those of combination with nickel or zinc. From those results, it could be suggested that the heavy metals decreased the viability of mouse fibroblast L929 cells in a concentration-dependent manner and have cytogenic toxic effects, but mixed group decreased the cytotoxic and cytogenic toxicity on L929 cells.

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Preparation and Dyeability of Reactive Dyes Fixable at Neutral pH (중성욕 고착형 반응성 염료의 제조 및 그의 염색성)

  • Choi, Chang Nam;Lee, Young Mi;Lee, Woong Eui
    • Textile Coloration and Finishing
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    • v.9 no.3
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    • pp.42-49
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    • 1997
  • In order to study the dyeability of reactive dyes fixable at neutral pH, three reactive dyes(DYE-NC, DYE-PC, and DYE-LC) were prepared by the reaction of Cibacron Brilliant Red 3B-A with nicotinic acid, pyridine, and lutidine, respectively. FT-IR and UV/Vis spectrophometry were used to identify the dyes prepared. While the maximum absorption wavelength (&{\lambda}_{max}&) of Cibacron Brilliant Red 3B-A was 515nm, the &{\lambda}_{max}& of DYE-NC, DYE-PC, and DYE-LC were 522nm, 525nm, and 536nm, respectively. The &{\lambda}_{max}& was shifted to the longer wavelength by introducing the electron donating groups to the pyridine ring of Cibacron Brilliant Red 3B-A. All of the reactive dyes synthesized showed good exhaustion and fixation property to cotton fabric at higher temperature and neutral pH condition. It was regarded that the quaternary pyridinium ion functionated as the leaving group instead of the chloride ion. Among them, DYE-NC showed the best dyeability at the above condition. It was considered that the electron withdrawing carboxylic acid group in nicotinic acid enhanced the cationic property of nitrogn in pyridine ring, resulting the good reactivity with OH group in cellulose.

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Evaluation of neutral red uptake assay using Balb/c 3T3 cells as a screening test to predict skin phototoxicity potential

  • Lee, Jong-Kwon;Lee, Eun-Hee;Kim, Ju-Hwan;Hong, Jin-Tae;Kim, Hyung-Soo;Park, Ki-Sook;Ahn, Kwang-Soo;Cho, Dae-Hyun;Lee, Sun-Hee
    • Proceedings of the Korean Society of Toxicology Conference
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    • 2001.05a
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    • pp.109-109
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    • 2001
  • In order to evaluate the neutral red uptake assay as an alternative method for phototoxicity test, we compared the potential of phototoxicity in vitro in cultured human fibroblasts and 3T3 fibroblast cells derived from Balb/c mice. Both fibroblasts were exposed to various known phototoxic chemicals (promethazine, neutral red, chlorpromazine, chlortetracycline, amiodarone, bithionol, 8-methoxypsoralen) and non-phototoxic chemical (ammonium laureth sulfate) and irradiated with 5 J/cm$^2$ of UVA.(omitted)

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Assessment of Immune Parameters of the Wild Pacific Oyster (Crassostrea gigas) using a Flow Cytometry and Neutral Red Retention Assay (유세포 분석기와 Neutral Red Retention Assay를 이용한 자연산 참굴(Crassostrea gigas)의 면역력 측정)

  • Hong, Hyun-Ki;Kang, Hyun-Sil;Kim, Young-Ok;Choi, Kwang-Sik
    • Ocean and Polar Research
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    • v.34 no.2
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    • pp.137-149
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    • 2012
  • Hemocyte parameters of the wild Pacific oyster Crassostrea gigas inhabiting intertidal zones in small bays (Gwangyang and Jinhae Bay) on the southern coast of Korea were evaluated using flow cytometry and neutral red retention (NRR) assay. Morphological features, cell count, mortality, DNA damage, phagocytosis, and lysosomal membrane stability of hemocytes were analyzed. Three types of hemocytes were identified in the oyster hemolymph: granulocytes, hyalinocytes, and blast-like cells. Immune related functions of hemocyte including phagocytosis and lysosomal membrane stability were significantly different among the study areas (P<0.05), while cell count, mortality, and DNA damage of hemocytes were not significantly different. In Gwangyang Bay, phagocytosis of granulocytes and lysosomal membrane stability of oyster hemocytes inhabiting inside bay were significantly lower than those of oyster hemocytes in outside bay (P<0.05), indicating that oysters in inside bay of Gwangyang were relatively suppressed the immunological function in hemocytes. Contrary to Gwangyang Bay, immune parameters of oyster hemocytes in Jinhae Bay not showed the difference between sampling sites. In conclusion, flow cytometry and NRR assay using oyster hemocyte has a powerful tool to investigate the cell level in a short time due to no-preprocessing of material.

Application of Neutral Red Staining Method to Distinguishing Live and Dead Marine Plankton for the Investigation of Efficacy of Ship's Ballast Water Treatment System (선박평형수 처리 시스템 효율 검증을 위한 해양 플랑크톤 생사판별시 Neutral red 염색법 적용 가능성 연구)

  • Hyun, Bonggil;Shin, Kyoungsoon;Chung, Hansik;Choi, Seo-Yeol;Jang, Min-Chul;Lee, Woo-Jin;Choi, Keun-Hyung
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.19 no.4
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    • pp.223-231
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    • 2014
  • In order to prevent the spread of non-indigenous aquatic species through the ballast water in commercial ships, International Maritime Organization (IMO) adopted in 2004 the International Convention for Control and Management of Ship's Ballast Water and Sediments. The Convention mandates treatment of ballast water for most transoceanic voyages and its confirmation of treatment is made with plankton live/dead assay. Fluorescein diacetate assay (FDA), which produces bright green light for live phytoplankton, has been a de facto standard method to determine the survival of marine plankton, but its staining efficacy has been in dispute. In the present study, we examined the limitation of FDA, and compared its efficacy with Neutral red (NR) staining, another promising assay and widely used especially for zooplankton mortality. For all phytoplankton species studied in the present study, except Ditylum brightwellii, the staining efficiency was <50% with FDA. The green FDA fluorescence interfered with phytoplankton autofluorescence in most samples. In contrast, NR assay stained over 90% of both phytoplankton and zooplankton species tested in this study. FDA assay also showed that green FDA fluorescence rapidly faded when phytoplankton cells were exposed to microscope light. Both FDA and NR assay were negative on formalin-killed individuals of both phytoplankton and zooplankton species. Our results suggest that NR assay is more effective for determining the survival of marine plankton and can be applied to test the efficacy of ballast water treatment.