• 제목/요약/키워드: nitrocellulose

검색결과 105건 처리시간 0.021초

Purification of Deoxycytidine Kinase from Various Human Leukemic Cells by End-product Analog Affinity Chromatography

  • Kim, Min-Young
    • BMB Reports
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    • 제28권4호
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    • pp.281-289
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    • 1995
  • Homogeneous human deoxycytidine kinase was purified in one step from a variety of spontaneous human leukemic cells (T-ALL, B-ALL, B-CLL, AML, CML), and from cultured T-lymphoblast cells (MOLT-4) using the newly developed affinity medium, $dCp_4$-Sepharose. Starting with an ammonium sulfate fraction, purification was achieved in one step with the kinase being eluted from a column by the end product inhibitor, dCTP. The purified deoxycytidine kinase from T-ALL cells phosphorylated deoxyadenosine and deoxyguanosine, as well as deoxycytidine. The enzyme purified from T-ALL and B-CLL cells yielded one major band with a molecular weight of 52 kDa determined by SDS-polyacrylamide gel electrophoresis. AML and CML cells yielded one 52 kDa band and an extra band of 30 kDa molecular weight. On the other hand, B-ALL and MOLT-4 cells showed a low molecular weight band of 30 kDa only. However, the electrophoretic mobilities of enzymatic activity in 12% non-denaturing gels were identical for the dCyd kinase from all different kinds of leukemic cell lines, except that the B-ALL, B-CLL, and MOLT-4 cell preparations had an extra minor peak, all at the same position. dAdo and dCyd phosphorylating activities comigrated indicating that these activities are all associated with the same protein. Two new methods, a disk implantation method and a nitrocellulose powder method were used with a small amount of enzyme protein to raise polyclonal antibodies against dCyd kinase purified from T-ALL cells.

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Monoclonal Antibody CFC-6, which Binds to Helix II, Inhibits Erythropoietin-Induced Bioactivity

  • Ha, Byung-Jhip;Kim, Suk-Joon;Park, Ji-Sook;Yoo, Ree-Ann;Lee, Dong-Eok;Yoo, Ook-Joon;Woo, Koo;Kim, Hyun-Su;Oh, Myung-Suk
    • BMB Reports
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    • 제30권5호
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    • pp.315-319
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    • 1997
  • It was discovered that monoclonal anti-erythropoietin (EPO) antibody CFC-6 can neutralize EPO-induced cell activation. To know the binding site of CFC-6, recombinant human erythropoietin (rhEPO) was digested with Glu-C, followed by a separation using high performance liquid chromato graphy (HPLC). Each HPLC fraction was blotted on the nitrocellulose membrane and the membrane was treated with anti-EPO antibody CFC-6 and anti-mouse antibody which is modified with peroxidase. Only one spot showed the color and the fraction was sequenced by Edman degradation. The results suggest that CFC-6 recognizes amino acid sequence V63-W-Q-G-L-A-L-L-S-E72 which is a part of helix II of the EPO molecule. Binding of CFC-6 to EPO may inhibit EPO binding to its receptor, which implies that the antibody binding site and the receptor binding site are close or overlapping.

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가소제를 포함한 셀룰로오즈 유도체의 유변학적 거동 (Rheological properties of cellulose derivative including plasticizer)

  • 최형진;유재림;김성태;현형수
    • 한국추진공학회:학술대회논문집
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    • 한국추진공학회 2005년도 제25회 추계학술대회논문집
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    • pp.9-12
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    • 2005
  • 본 연구에서는 셀룰로오즈 유도체로 사용되어지는 cellulose acetate butyate (CAB), cellulose acetate propionate (CAP), nitrocellilose (GC-519)의 분자량에 의한 유변학적 특성의 영향을 분석하기 위하여 각각의 셀룰로오즈 유도체를 Gel Permeartion Chromatography (GPC)를 이용하여 각각의 분자량을 측정하였다. GPC를 이용하여 셀룰로오즈 유도체의 중량평균분자량 (Mw)과 수평균분자량 (Mn)을 측정하였다. 각각의 셀룰로오즈 유도체에 가소제인 di-n-propyl adipate (DNPA)를 첨가한 뒤 아세톤에 녹여 레오미터를 이용하여 $0^{\circ}C$에서 가소제가 포함된 셀룰로오즈 유도체의 유변학적 특성을 측정하였다.

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일회용 스트립형 효소면역센서용 플랫폼의 개발 (Development of Disposable Enzyme-linked Immunosensor Strip Platform)

  • 최지혜;이승재;장승철;김경천
    • 센서학회지
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    • 제20권6호
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    • pp.400-405
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    • 2011
  • This study introduced the development of a strip type disposable enzyme-linked immunosensor platform for the detection of IgG. Strips of the strip sensor were fabricated by using commercial nitrocellulose filter membranes and a housing holder for the strips was manufactured by using a standard injection molding process for a plastic material. An IgG-urease conjugate was prepared and used for the competitive immune-binding with sample IgG. From the enzymatic reaction between the conjugated urease and urea added, ammonia was generated and caused a localized alkaline pH change on the immobilized antibody band which was coated onto the sensor strips. This pH increase subsequently caused a color change of the antibody band in the presence of a pH indicator, phenol red. Used in conjunction with a competitive immunoassay format, the intensity of the color produced is directly linked with the concentration of target analyte, IgG, and specific measurement of IgG in a lateral flow immunoassay format was achieved over the range 100 ppb to 2000 ppb IgG.

Early recognized antigen (p34) of Toxoplasma gondii after peroral ingestion of tissue cyst forming strain (Me49 strain) in mice

  • Park, Yun-Kyu;Nam, Ho-Woo
    • Parasites, Hosts and Diseases
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    • 제37권3호
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    • pp.157-162
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    • 1999
  • Serum from mouse orally ingested with tissue cyst forming stain (Me49) of Toxoplasma gondii was assayed by Western blot and immunofluorescene assay (IFA) to establish early responses in antigenicity of the parasite in mouse model of foodborne toxoplasmosis. Sera were collected weekly to blot the RH antigen transferred onto nitrocellulose paper after being separated by 12% SDS-PAGE. With the second week serum, 34 kDa protein (p34) was detected uniquely, and all antigens of T.gondii were detected with the sera from 3 or 4 weeks. p34 was not a member of the major surface membrane proteins and confirmed to be localized in the rhoptry by IFA. It was secreted into parasitophorous vacuolar membrane (PVM) during the entry into host cells. 10.3% of sera detected p34, while all the ELISA positive sera detected the band. It has diagnostic usefulness of presumed T.gondii infection. We suggest the name of the p34 protein as ROP9.

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The Distribution of Cytoplasm and Nuclei within the Extra-radical Mycelia in Glomus intraradices, a Species of Arbuscular Mycorrhizal Fungi

  • Lee, Jai-Koo
    • Mycobiology
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    • 제39권2호
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    • pp.79-84
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    • 2011
  • Nuclear distribution within the extra-radical fungal structures and during spore production in the arbuscular mycorrhizae fungus Glomus intraradices was examined using an in vitro monoxenic culture system. A di-compartmental monoxenic culture system was modified using a nitrocellulose membrane and a coverglass slip for detailed observations. Nuclear distribution was observed using the fluorescent DNA binding probes SYBR Green I and DAPI. Both septate and non-septate mycelial regions were observed, but cytoplasmic contents were only found within non-septate mycelia. Nuclear fluorescent staining revealed that the non-septate hyphal region contained nuclei only with cytoplasm, and that nuclear distribution was limited by septa. Swollen hyphal bodies were often associated with septate and empty-looking hyphae. Cytoplasmic contents filled the swollen hyphal body from the non-septate hyphal region following removal of the septa. As a consequence, the swollen body developed into a new spore. These observations provide understanding about the distribution of AM fungal nuclei within extra-radical mycelia and during spore formation. The results suggest a mechanism by which the development of a cytoplasm-containing mycelium is controlled by the formation or removal of septa to efficiently maintain and proliferate essential contents. This mechanism may provide a survival strategy to the fungus.

면역크로마토그라피 기법을 이용한 Salmonella 속균 신속 검출킷트 개발 (Development of the rapid detection kit for Salmonella spp. using immunochromatographic assay)

  • 정병열;정석찬
    • 대한수의학회지
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    • 제45권2호
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    • pp.191-197
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    • 2005
  • An immunochromatographic (IC) strip for the rapid detection of Salmonella spp. in the enriched sample was developed. Affinity purified Salmonella polyclonal antibody was conjugated with 40 nm colloidal gold particles which were prepared by citrate method in our laboratory. The antigen-antibody-gold complex was captured by Salmonella antibody attached to test line of nitrocellulose membrane during the capillary migration of sample. Specificity of the IC strip was calculated to be 100% (12/12) and sensitivity was 97.6% (41/42) in the test with pure cultured bacteria. Salmonella was artificially inoculated into raw pork macerated with enrichment broth. And then it was 10-fold diluted from $5.2{\times}10^{8}CFU/ml$ to 5.2 CFU/ml. The IC strip could detect $5.2{\times}10^{6}CFU/ml$ before enrichment. However, the lowest limit of detection was 5.2 CFU/ml after overnight incubation. The results indicated that the IC assay was a rapid, economical and simple method with high specificity and sensitivity for the detection of Salmonella spp. without using any equipment.

활성탄소섬유를 이용한 추진제 저장수명 연장 (Propellant Shelf-life Extension by Surface-modified Activated Carbon Fiber)

  • 윤근식;이영석;유승곤
    • Korean Chemical Engineering Research
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    • 제49권4호
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    • pp.443-448
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    • 2011
  • 추진제는 저장 중 발생되는 질소산화물로 인해서 저장수명이 짧아진다. 추진제의 저장수명을 연장할 목적으로 활성탄소섬유로 추진제에서 발생하는 질소산화물을 흡착하였다. 활성탄소섬유에 폐추진제를 첨착시키고 열처리하여 표면을 개질한 결과 비표면적이 약간 감소하였으나 피리딘(pyridine), 피리돈(pyridone) 및 피롤(pyrrol) 등의 질소기능기가 생기는 것을 확인하였다. NO에 대한 흡착시험을 통해서 표면개질한 활성탄소섬유의 흡착능이 개질 이전의 활성탄소섬유에 비해 약 2배 증가하였다. 그리고 추진제에 대한 가속수명시험 결과 표면개질한 활성탄소섬유를 동봉하면 추진제의 저장수명이 약 25% 증가하였다.

니트로글리세린의 이동과 안정제(DPA, ECL)에 의한 소진탄피 저장수명 연구 (The Study on the Shelf Life of the Combustible Cartridge Case by the Stabilizer(DPA, ECL) and Migration of Nitroglycerin)

  • 임호영;장일호;서지현;정용근;조민수;한창호
    • 한국군사과학기술학회지
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    • 제22권4호
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    • pp.509-516
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    • 2019
  • It is well known that nitroglycerin(NG) may evaporate and migrate from the triple base propellant grains in storage. This physical process makes it double base environment to the CCC(combustible cartridge case) which is based on nitrocellulose(NC) without NG. Meanwhile, it is not appropriate to use diphenylamine(DPA) as a stabilizer for CCC in this double base environment because of incompatibility between DPA and NG. So we estimated the shelf life to study the effect of NG migration from propellant to CCC by following the procedures in the STANAG 4257. And we found out that CCC with ethylcentralite(ECL) has 7.5 years longer shelf life than with DPA, when NG migrates to CCC from triple base propellant grains.

RDX를 적용한 다기추진제의 연소 및 강내탄도 특성 (Characteristic Property of Combustion and Internal Ballistics of Triple-Based Propellant including RDX)

  • 손수정;이원민;이우진;권순길;정진영
    • 한국군사과학기술학회지
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    • 제25권3호
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    • pp.321-328
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    • 2022
  • The current development tend of the gun propellants that they should have low sensitivity and high energy. We studied a nitrocellulose based propellant composition that replaced sensitive NG with RDX and DEGDN which high energy and low sensitivity. The important factors in the design of the gun propellant were impetus and flame temperature. NC-based propellant containing RDX showed similar impetus but low flame temperature compared to KM30A1, a triple-based propellant. The developed propellant composition didn't show any abnormal combustion reaction and the characteristics of ballistic resistance were also confirmed.