• Title/Summary/Keyword: Sephadex

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Purification of Radiolabeled RNA Using Sephadex G-15 or G-50 Chromatography (Sephadex G-15 또는 G-50 chromatography를 이용한 방사성 동위원소로 표지된 RNA의 정제)

  • Yoo, Beong-Gyu;Lee, Jong-Seok
    • Journal of radiological science and technology
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    • v.21 no.1
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    • pp.65-68
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    • 1998
  • We attempted to purify radiolabeled RNA using Sephadex G-15 and G-50 chromatography instead of commercial RNA purification kit. In the Sephadex G-15 chromatography the major portion of RNA was eluted in the fractions ranging from 3rd to 5th whereas broad elution profile of RNA was obtained from the Sephadex G-50 chromatography. The elution profile and purity of RNA obtained from Sephadex G-15 chromatography was very similar to that by commercial RNA purification kit. Furthermore, operating time required for purification of RNA by Sephadex G-15 was rather smaller than that by commercial kit. Overall results suggest that the purification of radiolabeled RNA using Sephadex G-15 is more money and time saying than using commercial RNA purification kit.

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Comparative Studies of Invertase Isozymes Produced by Rhodotorula glutinis K-24 (Rhodotorula glutinis K-24가 생산하는 Invertase Isozymes군에 관한 비교 연구)

  • Lee, Tae-Ho;Kim, Chul;Lee, Sang-Ok
    • Microbiology and Biotechnology Letters
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    • v.17 no.4
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    • pp.313-320
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    • 1989
  • Rhodotorula glutinis K-24 was found to produce internal, cell wall bound, and external invertase. Internal invertase was purified by column chromatographies on DEAE-Sephadex A-50, Sp-sephadex C-50, gel filtration on Sephadex G-200 and isoelectric focusing. Cell wall bound invertase was partially purified by the following procedures; column chromatography on DEAE-Sephadex A-50 and gel filtration on Sephadex G-100. Optimum pH and temperature for enzymatic activities of internal and cell wall bound invertase were pH 3.0 and 6$0^{\circ}C$, respectively. Both enzymes were inhibited by HgC1$_2$, AgNO$_3$, MnSO$_4$, and sodium dodecylsulfate. The molecular weights of internal and cell wall bound invertases were estimated to be 310,000 and 61,000, respectively. Other physicochemical properties of the both enzymes were similar.

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Effect of Saline-Sephadex on the Organ Distribution of $^{99m}TC$-Sephadex (식염수-Sephadex가 $^{99m}TC$-Sephadex의 폐분포에 미치는 영향)

  • Sung, Ho-Kyung;Kang, Sin-Koo;Koh, Joo-Hwan;Moon, Kwang-Nam;Lee, Jang-Kyu
    • The Korean Journal of Nuclear Medicine
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    • v.5 no.2
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    • pp.57-64
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    • 1971
  • The organ distribution study and the whole body scan were done in the albino rats at intervals of 5, 30, 60, 90 and 120 minutes after the intravenous injection of $^{99m}TC$-pertechnetate absorbed in Sephadex beads of $20{\sim}80$ micra in diameter. Effect of additional injection of physiological saline and saline absorbed in Sephadex beads of $20{\sim}80$ micra in diameter on the scan and organ distribution were also studied. The results were as follows. 1. Five minutes after the injection of $^{99m}TC$-pertechnetate absorbed in Sephadex beads of $20{\sim}80$ micra in diameter, Sephadex was well trapped in the lungs, with which the excellent lung scan was obtained. Two hours after the injection, kidneys were well visualized instead of lungs, which suggested that kidney acts as the excretory organ. Five minutes prior to scan, additional injection of physiological saline absorbed in Sephadex above mentioned was done. The bladder was also well visualized together with the kidneys. 2. In the distribution studies, most of radioactivity was detected in the lungs at 5 minutes and was gradually transferred chiefly to the kidneys and bladder and partly to the liver. 3. Additional injection of physiological saline resulted in a rapid transfer of $^{99m}TC$ trapped in the lung to both the kidneys and liver. 4. Additional injection of physiological saline absorbed in Sephadex beads of $20{\sim}80$ micra in diameter resulted in a rapid transfer of $^{99m}TC$ trapped in the lung to only the urinary system. 5. Results of these studies suggested that; a) Other nutrients and therapeutic compounds may be carried into the lungs along with Sephadex beads and then released in high concentration, which would exert greater therapeutic effect locally than that of the usual administration. b) Some radionuclides absorbed in Sephadex could be used as the lung scan agents, the flushing out of which by Sephadex-saline also give satisfactory renal and bladder scans. c) Other potent therapeutic radionuclides could be retained for some time by this method, which can be in the lungs easily flushed out within 2 hours.

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Radius and Length of Sephadex G Gel Fibers (Sephadex G Gel 섬유의 반경과 길이)

  • 구윤모
    • KSBB Journal
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    • v.4 no.1
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    • pp.15-16
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    • 1989
  • The dimensions of dextran gel fiber described by the Ogston-Laurent gel model were estimated for Sephadex G gels. The length of fibers are ranging from $12.48{\times}10^{12}(G-25)\;to\;1.76{\tiems}10^{12}(G-200)$ cm/ml. The radius of gel fiber and the partial specific volume of dextran gel are $7.21{\tiems}10^{-8}$ cm and 0.586 ml/g, respectively.

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Evaluation of Extended Canine Semen after Different Filtration Treatment (개 희석 정액의 다양한 filtration 처리 후 정자평가)

  • Kim, Yong-Jun;Kim, Jin-Young;Kim, Sue-Hee;Lee, Young-Jun
    • Journal of Veterinary Clinics
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    • v.24 no.4
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    • pp.577-583
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    • 2007
  • It is important to obtain semen with good quality for efficient fertilization and pregnancy. To obtain these semen, various methods have been developed but most of these methods are time consuming and require costly equipment. Therefore, the objective of this research is to investigate the usability of column filtration system as quick and simple method to get sperm with better quality. Ejaculates were obtained from 5 dogs and analyzed with basic quality parameters before each filtration. Sperm concentration was adjusted to $5{\times}10^7/ml$ after dilution. The experimental groups were divided into non-filtered group(control) and filtered groups(glass wool, Sephadex 5% and Sephadex 20%). Ejaculates were filtered through each filter system and assessed by recovery rate of sperm, motility, normal morphology, CFDA/PI stain and plasma membrane integrity(hypo-osmotic swelling test, HOST). The lowest recovery rate of spermatozoa was recorded in glass wool filtration group, followed by 20% Sephadex filtration group(p<0.05). There was no significant difference between control(non-filtered) and 5% Sephadex filtration poop. Also, there was no significant difference of sperm motility assessed under light microscope among experimental groups. Morphological normality of canine spermatozoa was the highest in the glass wool filtration group and the lowest in the 5% Sephadex filtration group with no significant differences versus 20% Sephadex filtration and control group, respectively(p<0.05). Viability of canine sperm assessed by CFCA/PI staining was the highest in the glass wool filtration poop with no significant difference versus the control group, and the lowest in the 20% Sephadex filtration group with no significant difference versus 5% Sephadex filtration group, respectively(p<0.05). HOS values of canine sperm was the highest in the 20% Sephadex filtration group with no significant difference versus 5% Sephadex filtration group, and the lowest in the control poop with no significant difference versus glass wool filtration group, respectively(p<0.05). Therefore, these results indicated that filtration treatment for extended canine sperm would be useful method to get sperm with better quality by trapping the damaged sperm, consequently filter would be physical barrier against injured or immotile sperm.

Purification and Properties of an Extracellular Chitinase from Streptomyces sp. (Streptomyces속 균주로 부터 생산되는 Chitinase의 정제 및 그 성질)

  • Hong, Yong-Ki;Seu, Jung-Hwn
    • Microbiology and Biotechnology Letters
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    • v.7 no.3
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    • pp.149-155
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    • 1979
  • Streptomyces sp. 115-5 was selected as the most active microorganism of about 200 strains for the production of chitinase. The enzyme was purified by (NH$_4$)$_2$SO$_4$ treatment, 1st-Sephadex G-100, DEAE-Cellulose, 2nd-Sephadex G-100 column chromatography, and evidence for homogenity was obtained from CM-Sephadex C-50 column chromatography and polyacylamide gel electrophoresis. The purified enzyme hydrolyzed chitin (N-acetyl glucosamine polymer) and chitosan (glucosamine polymer) but not cellulose. And with chitin as the substrate, a Km value of 3.6 mg of chitin per ml and a Vmax of 100 $\mu$mo1e fer hr were found. The activation of the chitinase was 3.66 kcal per mole. The molecular weight of the enzyme was esti-mated about 56,000 daltons by Sephadex G-100 chromatography and isoelectric point as pH 3.0.

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The Effects of Fractions of Chick Embryo Extract on the Fusion of Cultured Chick Embryonic Myoblasts (培養 鷄胚 筋原細胞의 融合에 미치는 鷄胚 抽出液 分劃의 영향)

  • Ha, Doo-Bong;Lee, Chung-Choo;Park, Young-Chul;Lim, Woon-Ki;Yoo, Byoung-Je
    • The Korean Journal of Zoology
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    • v.28 no.3
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    • pp.179-193
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    • 1985
  • In order to find factors which are essential for the differentiation of chick embryonic myoblasts in culture, chick embryo extract was fractionated by ammonium sulfate or/and Sephadex G-75, and the effects of each fraction on the proliferation and fusion of the myoblasts were examined. The results obtained were as follows: (1) High concentration of embryo extract in the culture medium enhanced the cell proliferation and delayed the fusion of myoblasts. (2) The Sephadex G-75 fractions of embryo extract having proteins of molecular weight between 40,000 and 22,000 enhanced the proliferation and fusion of myoblasts when added to culture media. (3) The fraction of embryo extract precipitated in $60\\sim95%$ saturated ammonium sulfate solution enhanced evidently both the proliferation and fusion of myoblasts. Elution of this effective fraction by Sephadex G-75 showed similar elution profile and effects on the myoblast differentiation as those observed by Sephadex G-75 chromatography of the whole embryo extract, suggesting that the Sephadex fractions and ammonium sulfate fractions contain the same factors that enhance the proliferation and fusion of myoblasts.

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Quality and Fertility of Post Thaw Sephadex Filtered Bull Semen

  • Kumar, A.;Singh, J.;Nanda, A.S.;Pangaonkar, G.R.
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.6
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    • pp.755-759
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    • 2004
  • The present investigation was carried out to assess the effect of Sephadex (G-15) filtration on the post thaw bull semen quality and conception rate. Post thaw unfiltered (control) and Sephadex filtered semen from four healthy bulls (three cross bred and one pure bred Holstein Friesian) were subjected to microscopic examination viz. sperm concentration, individual motility, live sperm count and sperm morphology. Sixty-two healthy, normal cycling crossbred cows were inseminated with post thaw unfiltered (n=32) and filtered semen (n=30). Sephadex filtration of post thaw semen significantly (p<0.05) decreased total sperm concentration and sperm with abnormal head, mid piece and tail. The overall average total sperm concentration, head and tail defects in filtered semen decreased significantly (53.4, 1.2 and 6.4 million) than in the unfiltered semen (80.4, 2.4 and 15.7 million, respectively). However, after filtration significant (p<0.05) increase in overall average motile and live sperm concentration were observed (38.8 and 38.0) as compared to unfiltered semen (29.2 and 32.0 million, respectively). The overall conception rate recorded was 21.9% with post thaw unfiltered semen and 56.7% with filtered semen. It was concluded that Sephadex filtration of post thaw semen improved its quality and conception rate.

Substrate Specificity of Cellulase from Aspergillus niger (Aspergillus niger가 생산하는 섬유소 분해효소의 기질에 대한 특이성)

  • Oh, Tae-Kwang;Park, Kwan-Hwa;Shin, Hyun-Kyung;Kim, Ze-Uook
    • Applied Biological Chemistry
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    • v.28 no.3
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    • pp.162-166
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    • 1985
  • Three isozymes of Carboxymethyl Cellulase $(FI^*,\;FII^*,\;FIII)$ and two fractious of ${\beta}-1,4-D-Cellobiohydrolase$(CI, CIl) from Aspergillus niger were purified by Sephadex G-150, DEAE-Sephadex and Sephadex G-75 column chromatography. From the results of enzymatic hydrolysis and X-ray diffraction, ${\beta}-1,4-D-Cellobiohyarolase$ has a high activity toward highly crystalline cellulose such as filter paper and acts synergistically with Cx enzyme.

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