• Title/Summary/Keyword: Nitrocellulose

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

  • Kim, Min-Young
    • BMB Reports
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    • v.28 no.4
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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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    • v.30 no.5
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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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Rheological properties of cellulose derivative including plasticizer (가소제를 포함한 셀룰로오즈 유도체의 유변학적 거동)

  • Choi Hyoung-Jin;You Jae-Lim;Kim Sung-Thae;Hyun Hyoung-Soo
    • Proceedings of the Korean Society of Propulsion Engineers Conference
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    • 2005.11a
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    • pp.9-12
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    • 2005
  • Molecular characteristics and rheological properties of cellulose acetate butyrate (CAB), cellulose acetate propionate (CAP) and nitrocellulose (GC-519) which are being widely used as propellants were investigated. Their weight-average molecular weight (Mw) and number-average molecular weight (Mn) were estimated via Gel Permeartion Chromatograpy (GPC). Cellulose derivatices were mixed with di-n-propyl adipate (DNPA) which acted as plasticizer in acetone, and then rheological properties of the mixture of cellulose derivatives and this plasticizer in acetone were investigated at $0^{\circ}C$ by rheometer.

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

  • Choi, Ji-Hye;Yi, Seung-Jae;Chang, Seung-Cheol;Kim, Kyung-Chun
    • Journal of Sensor Science and Technology
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    • v.20 no.6
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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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    • v.37 no.3
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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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    • v.39 no.2
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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.

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

  • Jung, Byeong-yeal;Jung, Suk-chan
    • Korean Journal of Veterinary Research
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    • v.45 no.2
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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.

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

  • Yoon, Keun Sig;Lee, Young Seak;Ryu, Seung Kon
    • Korean Chemical Engineering Research
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    • v.49 no.4
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    • pp.443-448
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    • 2011
  • The propellant has a short shelf-life because of nitrogen oxides that were released from nitrocellulose decomposition. As-received and surface-modified ACFs were applied to remove the nitrogen oxides with intend to extend the shelf-life of propellant. The specific surface area of modified ACFs was slightly decreased but nitrogen function groups such as pyridine, pyridone and pyrrol were created on the surface of ACFs. As a result, the NO removal capacity of the surface-modified ACF by propellant waste increased about twice than that of the as-received ACF. The shelf-life of propellant was extended about 1.25 times by accompanying surface-modified ACF.

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

  • Lim, Hoyoung;Jang, Ilho;Seo, Jihyun;Jung, Yonggeun;Jo, Minsoo;Han, Changho
    • Journal of the Korea Institute of Military Science and Technology
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    • v.22 no.4
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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.

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

  • Son, Soojung;Lee, Wonmin;Lee, Woojin;Kwon, Soonkil;Jung, Jinyoung
    • Journal of the Korea Institute of Military Science and Technology
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    • v.25 no.3
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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.