• Title/Summary/Keyword: Silver staining

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Dye-silver double staining method for proteins in SDS-polyacrylamide gels using a dye as a silver sensitizer

  • Jin, Li-Tai;Hwang, Sun-Young;Yoo, Gyurng-Soo;Choi, Jung-Kap
    • Proceedings of the PSK Conference
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    • 2003.10b
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    • pp.145.2-145.2
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    • 2003
  • We have developed a silver staining method using a dye as a silver sensitizer. Dye staining is performed in combination with silver nitrate staining. Dye-silver staining shortens the time of silver staining (~1 hr) and improves the sensitivity better than that of silver diamine stain (1-10 ng) or comparable to that of silver nitrate stain with glutaraldehyde as a silver sensitizer. In dye staining (silver sensitizing step), it has been proven that the sensitivity is at least 4 times comparing with that of CBBR stain and staining time is about 45 min. (omitted)

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Cytogenetic Analysis of All-Female Triploid Olive Flounder Paralichthys olivaceus for Ploidy Verification (전 암컷 3배체 넙치(Paralichthys olivaceus)에 대한 효율적인 세포유전학 분석법)

  • Ko, Min Gyun;Jung, Hyo Sun;Lee, Hyo Bin;Kim, Dong Soo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.49 no.5
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    • pp.671-674
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    • 2016
  • We cytogenetically analyzed a triploid King-Nupchi strain of the olive flounder Paralichthys olivaceus to define the simplest, most rapid, and most effective method of ploidy analysis in aquaculture farms. Female triploidy of the flounder King-Nupchi strain was induced by cold shock (3 min post-fertilization at 2-4℃ for 45 min). Triploid induction was confirmed by erythrocyte measurement (nuclear volume, 29.15±2.10 μm3); flow cytometry (2.14±0.03 pg/cell); chromosome count (3N=72); Ag-NOR banding; and silver staining. Silver staining of finned cells obtained using a solid tissue technique was the most effective method of ploidy verification.

Further Purification of Radioprotective Ginseng Protein Fraction by Gel Filtration (Gel filtration에 의한 한방사선 인삼단백 분획의 정제)

  • 김춘미;박경애
    • Journal of Ginseng Research
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    • v.13 no.2
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    • pp.254-259
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    • 1989
  • A radioprotective ginseng protein fraction was obtained from Korean white ginseng powder by the following isolation and purification procedures: Tris-HCI buffer extraction, 70% ammonium sulfate fractionation, CM-rellulosr column chromatography, heat inactivation and Sephadex G-75 column chromatography. This fraction was further purified by Sepharose 4B and Sephadex G-150 column chromatographies. Three fractions obtained were subjected to Native-PAGE and SDS-PAGE using gradient gels and the silver staining method. Molecular weights of the native proteins and their subunits were estimated.

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Comparison of different ploidy detection methods in Oncorhynchus mykiss, the rainbow trout

  • Kim, Hong Seab;Chung, Ki-Hwa;Son, Jung-Ho
    • Fisheries and Aquatic Sciences
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    • v.20 no.11
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    • pp.29.1-29.7
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    • 2017
  • The objective of this study was to determine a simple and reliable ploidy identification protocol for the rainbow trout (RT), Oncorhynchus mykiss, in the field condition. To evaluate the ploidy level and compare different detection protocols, triploid RT and gynogenesis were induced by UV irradiation and/or heat shock. The hatching rate at day 30 was 85.2% and the survival rate at day 90 was 69.4% (fingerling). The sex ratio of female RT was 93.75% in the gynogenesis group, illustrating that the UV irradiation inactivated the sperm DNA. The hatching rate and survival rate were 82.0 and 74.7%, respectively, in the triploid-induced group. The triploid induction rate by heat shock procedure was 73.9%. Cytogenetic protocols for ploidy identification such as chromosome counting, erythrocyte nuclear size comparison, and analysis of nucleolar organizing regions (NORs) by silver staining were compared. Silver nitrate staining showed the greatest success rate (22/23 and 32/32 for the triploid-induced group and gynogenesis group, respectively), followed by erythrocyte nuclear size comparison (16/23 and 19/32 for the triploid-induced group and gynogenesis group, respectively) and, lastly, chromosome preparation (2/23 and 6/32 for the triploid-induced group and gynogenesis group, respectively) with the lowest success rate. Based on our findings, silver staining for RT ploidy identification is speculated to be highly applicable in a wide range of research conditions, due to its cost-effectiveness and simplicity compared to other numerous ploidy detection protocols.

The Color Fading and Staining of Fabrics by Drum-type Washer (드럼세탁기 사용시 세탁물의 변.퇴색 방지에 관한 연구)

  • Ryu, Hyo-Seon;Kim, Eun-Ah;Yun, Chang-Sang
    • Journal of the Korean Society of Clothing and Textiles
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    • v.32 no.6
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    • pp.947-958
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    • 2008
  • To study the effect of a washing machine with silver nano technology on its detergency, the discoloring of the dyed clothes and the staining of standards adjacent fabrics were examined. As the laundry specimen, cotton fabric dyed with reactive dyes and Polyester fabric dyed with disperse dyes were chosen; and as the adjacent fabrics, undyed cotton. polyester and nylon fabrics were chosen. The colorfastness was evaluated after washing under conditions that those washing temperature, liquor ratio, detergency concentration and the type of water were varied. When the clothes were washed with the tap water contains silver ion, the deposition of silver compounds into the washed clothes was measured. As a results, after the washing in the various conditions, discoloring of the dyed clothes was not intense. The higher the washing temperature and the lower the liquor ratio, the larger the staining appeared on the white fabrics; especially for the white nylon fabrics. The concentration of detergent and the type of water affected hardly the colorfastness. After the repeated washing with the water contains silver, whiteness of the cotton and the nylon fabrics were lower than the result after the washing with the tap water, and a quantity of silver ions was found on the washed clothes.

New record of five Euplotes species(Protozoa, Ciliophora) collected from South Korea

  • Jeong Hyeon Yeo;Pablo Quintela-Alonso;Jae-Ho Jung
    • Journal of Species Research
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    • v.12 no.3
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    • pp.203-211
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    • 2023
  • Five ciliate species of Euplotes were isolated from fresh and coastal water during a sampling survey to identify unrecorded ciliates in South Korea. Their morphology was investigated using live observation, protargol and "wet" silver nitrate staining methods. Brief descriptions and microphotographs of each species and a comparison with related species are provided. Euplotes focardii is characterized by an average size of 65×47 ㎛ after protargol impregnation, 6 dorsal and 3 ventral ridges and dorsal argyrome pattern of double-eurystomus type. Euplotes nobilii shows an average size of 34×20 ㎛ after protargol staining, 6 dorsal and 3 ventral ridges and dorsal argyrome pattern of double-patella type. Euplotes octocarinatus, the only freshwater species described in the present study, is characterized by an average size of 66×46 ㎛ after protargol impregnation, 6 dorsal and 3 ventral ridges and dorsal argyrome pattern of double-patella type. Euplotes petzi has an average size of 43×30 ㎛ after protargol staining, a macronucleus hook-shaped and dorsal argyrome pattern in double-patella type. Euplotes raikovi is characterized by an average size of 40×24 ㎛ after protargol staining, 6 dorsal and 3 ventral ridges and dorsal argyrome pattern of double-patella type.

Pneumocystis carinii pneumonia in pigs (돼지의 Pneumocystis carinii 폐렴 증례)

  • Jung, Ji-Youl;Kim, Ki-Seung;Kim, Dae-Yong;Kim, Jae-Hoon
    • Korean Journal of Veterinary Research
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    • v.47 no.3
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    • pp.321-324
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    • 2007
  • Pneumocystis (P.) carinii is an opportunistic fungal pathogen of many animal species and human, which can cause fatal pneumonia in immunocompromised individuals. Three 100-day-old pigs with progressive atrophy, anorexia and respiratory distress were submitted to the Cheju National University for diagnosis. Grossly, the lungs were enlarged with rubbery consistency. Histopathologically, the lungs were characterized by diffuse interstitial pneumonia with thickening of alveolar septa due to infiltration of macrophages and lymphocytes. Alveolar lumens were filled with a foamy eosinophilic proteinaceous material in which numerous punctiform organisms. The organisms were demonstrated as P. carinii by Grocott-methenamine-silver staining and immunohistochemistry in lungs of two pigs. In our best knowledge, this is believed to be the first report of P. carinii pneumonia in pigs in Korea.

Effects on the tissue reaction using compomer & Ketac Silver in the maxillary furcation in the beagle dogs (Compomer와 Ketac Silver로 성견 상악 이개부 병소 충전시 조직반응에 미치는 영향)

  • Ryu, Jea-Youn;Lim, Sung-Bin;Chung, Chin-Hyung;Lee, Chong-Heon
    • Journal of Periodontal and Implant Science
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    • v.33 no.4
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    • pp.705-715
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    • 2003
  • Procedures for treatment of molar furcation invasion defects range from open flap debridement, apically repositioned flap surgery, hemisection, tunneling or extraction, to regenerative therapies using bone grafting or guided tissue regenerative therapy, or a combination of both. Several clinical evaluations using regenerative techniques have reported the potential for osseous repair of treated furcation invasions. Regenerative treatment of maxillary molars are more difficult due to the multiple root anatomy and multiple furcation entrances therefore, purpose of this study was to evaluated histologically compomer and Ketac Silver as a barrier in the treatment of a bi-furcated maxillary premolar. Five adult beagle dogs were used in this experiment. With intrasulcular and crestal incision, mucoperiostcal flap was elevated. Following decortication with 1/2 high speed round bur, furcation defect was made on maxillary premolar. 2 month later one premolar was filled with compomer and the other premolar was filled with Ketac Silver. After 4, 8 weeks, the animals were sacrificed by vascular perfusion. Tissue block was excised including the tooth and prepared for light microscope with H-E staining. Results were as follows. 1. Compomer & Ketac Silver restoration were encapsulated fine connective tissue. 2. In 4 weeks, compomer & Ketac Silver restoration slightly infiltrated inflammatory cells but not disturb the new bone or new cementum formation. 3. In 8 weeks, compomer & Ketac Silver restoration were less infiltrated iflammatory cell and encapsulated fine connective tissue. 4. Therefore, compomer & Ketac Silver filling to the grade III maxillary furcations with multiple root anatomy and multiple furcation entrances is possible clinical method and this technique is useful method for maxillary furcation involvement but it is thought that periodic maintenance should be needed

Detection of Fragment Length Polymorphism of the VNTR Loci D1S80 and D2S123 by PCR Amplification, PAGE and Silver Staining

  • Nam, Hyun-Suk;Kim, Eun-Hee;Yoon, Wan-Hee;Lee, Kong-Joo
    • BMB Reports
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    • v.28 no.4
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    • pp.359-362
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    • 1995
  • The highly polymorphic variable number of tandem repeat (VNTR) loci in the human genome are informative markers for the genetic characterization of individuals in the paternity test and forensic science as well as for the study of human disease. In this study, VNTR loci D1S80 and D2S123 have been amplified by PCR and the amplified length polymorphic alleles were detected with a discontinuous vertical PAGE system and silver staining. For explicit DNA typing, PCR optimization, in which amplification efficiencies are similar over a wide range of allele sizes, non-specific amplifications are minimal, and new longer alleles have high amplification efficiency, has been performed by changing the PCR reaction buffer composition and thermal cycling conditions. It turned out that adding an appropriate amount of Tween 20 and NP40 to the PCR reaction buffer and raising the annealing temperature to $68^{\circ}C$ in thermal cycling made it possible for optimal VNTR loci amplification. A modified PAGE system for VNTR separation was established. Under these conditions, new longer alleles in the 01580 locus were discovered and 025123 pattern changes in colorectal tumors were observed. These technical tips are valuable for detecting various amplified fragment length polymorphisms.

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Rapid Detection of Fluorescent DNA on Denaturing Polyacrylamide Gel by Using Gel Scanner (겔스캐너를 이용한 변성아크릴아마이드 겔의 형광 DNA 검출)

  • Ku Ja-Hwan;Jeong Ji-Ung;Cho Young-Chan
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.50 no.spc1
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    • pp.228-230
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    • 2005
  • The denature polyacrylamide gel stain silver nitrate is used for routine nucleic acid detection. More sensitive stains, such as Vistra Green, SYBR Green are available to address a broad range of DNA applications requiring lower detection limits in polyacrylamide gel formats. Gel Scanners, laser-scanning instruments, provide sensitive fluorescence detection of DNA gel stains. We established one step fluorescent impregnation enhanced sensitivity with simple, rapid and low cost. We have applied this fluorescent staining procedure for the routine analysis of DNA profiles generated by SSR amplification.