• Title/Summary/Keyword: silver staining method

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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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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

Improvement of micronucleus assay in the lymphocytes using Argyrophilic nucleolor organizer region(AgNOR) staining (Argyrophilic nucleolar organizer region(AgNOR) 염색을 이용한 림프구의 미세핵분석법의 개선)

  • Kim, Sung-ho;Han, Dong-un;Kang, Mun-il;Lim, Jeong-taek
    • Korean Journal of Veterinary Research
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    • v.36 no.2
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    • pp.277-281
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    • 1996
  • A technique to improve the analysis of micronuclei(MN) in lymphocytes as a cytogenetic indicator is reported. For the purpose of diminishing the variation of the result from individual reader and making it easier to distinguish accurately a cytokinesis blicked(CB) lymphocyte and micronuclei, we tried a modified one-step silver staining technique as a method for detection of the argyrophilic nucleolar organizer region(AgNOR) with or without conventional Giemsa stain in the slide from CB method. Compared with the conventional Giemsa stain, the preparation processed with this method are especially useful for the accurate analysis of MN of cultured lymphocyte with cytochalasin B. This method will be a useful technique for automated calculation of MN.

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Isolation and Purification of Lipopolysaccharide Derived from Escherichia coli O157:H7 for the Specific Antibody Production (병원성 Escherichia coli O157:H7의 특이 항체 생산을 위한 Lipopolysaccharide분리 및 정제)

  • 최학선;신영민;정숙현;박영민;안원근
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.3
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    • pp.571-575
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    • 2004
  • Escherichia coli O157:H7 cause hemorrhagic colitis and the extraintestinal complication of hemolytic-uremic syndrome, with their higher incidence occurring in children. Lipopolysaccharide (LPS) of E. coli O157:H7 is very important to make IgG anti-LPS with bactericidal activity. To identify the characteristic of E. coli OI57:H7, we isolated 60 MDa plasmid and amplified stx genes of shiga-like toxin (Stx) 1, 2 of E. coli O157:H7 by polymerase chain reaction (PCR) method. Using the simple purification method which contained phenol extract, ethanol precipitation and gel filtration steps, the LPS of E. coli O157:H7 was isolated and purified. Finally, we confirmed the purity of LPS through SDS-PAGE and silver nitrate staining.

Paternity test in dogs by microsatellite allele analysis (Microsatellite 대립유전자 분석을 통한 개에서의 친자감별)

  • Chae, Young-jin;Kim, Dong-keon;Kim, Hana;Lee, Moon-han;Hwang, Woo-suk;Lee, Byoung-chun;Youn, Hwa-young;Lee, Hang
    • Korean Journal of Veterinary Research
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    • v.39 no.1
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    • pp.213-219
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    • 1999
  • Microsatellite allele analysis has been used for individual identification and paternity test. In the present study, the biological father of three puppies was determined by using microsatellite allele amplification analysis. The mother bitch of the litter was a Poongsan dog. The three stud dogs that could have inseminated the bitch, by being in the same residence, were a white Poosan dog, a mixed breed, and a white Jindo dog. DNA was obtained from all the relevant dogs by buccal swabbing. Four loci of tetranucleotide repeat microsatellite were PCR-amplified, and analyzed by polyacrylamide gel electrophoresis and silver staining. The results of genotyping unambigously assigned the Poongsan dog as the biological father. There was no evidence of superfecundation. Therefore, the present study demonstrated the usefulness of microsatellite allele analysis as a simple, efficient method of paternity test in dogs.

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Glycogen distribution of germ cells and Sertoli cells of seminiferous tubules in Jindo dog (진도견 정세관의 정세포와 Sertoli 세포내 glycogen의 분포)

  • Park, Young-seok;Lee, Seong-ho
    • Korean Journal of Veterinary Research
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    • v.36 no.3
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    • pp.521-529
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    • 1996
  • In an effort to obtain basic data of carbohydrate metabolism during spermiogenesis of the sexually-matured Jindo dog, the glycogen distribution in the testis was investigated by light and transmission electron microscopy. Periodic acid thiocarbohydrazide silver proteinate physical development(PA-TCH-SP-PD) staining method provided better results in the detection of glycogen granules from Sertoli cells and germ cells than the periodic acid schiff(PAS) staining method did. Pre-treatment of the tissue sections with ${\alpha}$-amylase elicited a significant decrease in PA-TCH-SP-PD stained granules, which suggested that the stained granules were of glycogen origin. High concentration of the glycogen granules were observed in the Sertoli cells, especially in its column, sheet-like processes, club-like processes, and tubular processes. The glycogen granules were unevenly distributed in some Sertoli cell columns. These results strongly indicated that the Sertoli cells of Jindo dogs showed vigorous activity of carbohydrate metabolism.

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Paternity test in dogs by DNA analysis (유전자감식에 의한 개에서의 친자감별)

  • Lee, Hang;Chae, Young-Jin;Lee, Byeong-Chun
    • Journal of Veterinary Clinics
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    • v.15 no.2
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    • pp.274-278
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    • 1998
  • The biological father of two Golden Retriever puppies was determined between two proposed stud dogs by using microsatellite DNA analysis. DNA was obtained from all the relevant dogs by buccal swabbing and three loci of tetranucleotide repeat microsatellite were PCR-amplifiedl and analyzed by polyacrylamide gel electrophoresis and silver staining. One of the two proposed stud dogs was assigned as the biological father of the puppies by the genotyping. The result demonstrated that the microsatellite DNA analysis is a simple, efficient method of paternity test in dogs.

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The Significance of AgNOR Count in Body Fluid - Differential between reactive mesothelial cells & malignant cells - (체액도말에서의 AgNOR수의 유의성 - 반응성 중피세포와 악성세포의 감별 -)

  • Paik, Seung-Sam;Hong, Eun-Kyung;Jang, Se-Jin;Park, Moon-Hyang;Lee, Jung-Dal
    • The Korean Journal of Cytopathology
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    • v.8 no.2
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    • pp.129-134
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    • 1997
  • To distinguish reactive mesothelial cells from malignant cells in body fluid, we applied silver staining of nucleolar organizer regions(AgNORs) to ethanol fixed cytologic preparations. Fifty aspirated samples of benign(22 cases) and malignant(26 cases) body fluids were studied using the one step silver staining method. Two cytologically atypical samples were also included in the study. In malignant cases the mean AgNOR count was $3.56{\pm}0.81$, while in benign cases the mean AgNOR count was $2.02{\pm}0.33$. The difference of AgNOR counts between these two groups were statistically significant(p<0.001). The mean of atypical cases was 2.91. Both were diagnosed as malignant in follow-up cytology. In malignant effusions, there is statistically significant difference in AgNOR counts between cells forming complex papillae or clusters and singly scattered cells(p<0.05), $3.29{\pm}0.95\;and\;3.83{\pm}0.55$, respectively. We concluded that AgNOR count appears to be useful as a diagnostic tool especially when the cytologic differentiation is difficult.

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