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Analytical biochemistry 13건

  1. [해외논문]   Editorial Board   SCI SCIE


    Analytical biochemistry v.536 ,pp. ii - iii , 2017 , 0003-2697 ,

    초록

    원문보기

    원문보기
    무료다운로드 유료다운로드

    회원님의 원문열람 권한에 따라 열람이 불가능 할 수 있으며 권한이 없는 경우 해당 사이트의 정책에 따라 회원가입 및 유료구매가 필요할 수 있습니다.이동하는 사이트에서의 모든 정보이용은 NDSL과 무관합니다.

    NDSL에서는 해당 원문을 복사서비스하고 있습니다. 아래의 원문복사신청 또는 장바구니담기를 통하여 원문복사서비스 이용이 가능합니다.

    이미지

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  2. [해외논문]   Isolation of mouse chromaffin secretory vesicles and their division into 12 fractions   SCI SCIE

    Pardo, Marta R. (Corresponding author. Unidad de Farmacología, Facultad de Medicina, Universidad de La Laguna, E-38320 La Laguna, Tenerife, Spain.) , Esté , vez-Herrera, Judith , Castañ , eyra, Leandro , Borges, Ricardo , Machado, José , David
    Analytical biochemistry v.536 ,pp. 1 - 7 , 2017 , 0003-2697 ,

    초록

    Abstract The study of chromaffin secretory vesicles (SVs) has contributed immensely to our understanding of exocytosis. These organelles, also called chromaffin granules, are a specific type of large dense secretory vesicle found in many endocrine cells and neurons. Traditionally, they have been isolated from bovine adrenal glands due to the large number of SVs that can be obtained from this tissue. However, technical advances now make it possible to obtain very pure preparations of SVs from mice, which is particular interesting for functional studies given the availability of different genetically modified strains of mice. Despite the small size of the mouse adrenal medulla (400–500 μm and less than 2 mg in weight), we have successfully carried out functional studies on SVs isolated from WT and knockout mice. As such, we present here our method to purify crude vesicles and to fractionate mouse chromaffin SVs, along with examples of their functional characterization. Highlights We present a method to obtain high-yields of purified secretory vesicles from mouse chromaffin cells under isotonic conditions. We describe their further separation into 12 fractions and the analysis of the content of each fraction. These isolated vesicles retain the ability to take up catecholamines.

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    무료다운로드 유료다운로드

    회원님의 원문열람 권한에 따라 열람이 불가능 할 수 있으며 권한이 없는 경우 해당 사이트의 정책에 따라 회원가입 및 유료구매가 필요할 수 있습니다.이동하는 사이트에서의 모든 정보이용은 NDSL과 무관합니다.

    NDSL에서는 해당 원문을 복사서비스하고 있습니다. 아래의 원문복사신청 또는 장바구니담기를 통하여 원문복사서비스 이용이 가능합니다.

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    Fig. 1 이미지
  3. [해외논문]   A method for extracting and characterizing RNA from urine: For downstream PCR and RNAseq analysis   SCI SCIE

    Zhou, Kun (University of Minnesota Medical School –) , Spillman, Monique A. (Duluth, 1035 University Drive, Duluth, MN 55812-3031, USA ) , Behbakht, Kian (Texas A&M University Medical School, Baylor University Medical Center, Dallas, TX, USA ) , Komatsu, Julia M. (University of Colorado Anschutz Medical Campus, Aurora, CO 80045, USA ) , Abrahante, Juan E. (University of Minnesota Medical School –) , Hicks, Douglas (Duluth, 1035 University Drive, Duluth, MN 55812-3031, USA ) , Schotl, Brent (University of Minnesota Informatics Institute, University of Minnesota, Minneapolis, MN, USA ) , Odean, Evan (University of Colorado Anschutz Medical Campus, Aurora, CO 80045, USA ) , Jones, Kenneth L. (University of Minnesota Medical School –) , Graner, Michael W. (Duluth, 1035 University Drive, Duluth, MN 55812-3031, USA ) , Bemis, Lynne T. (University of Minnesota Medical School –)
    Analytical biochemistry v.536 ,pp. 8 - 15 , 2017 , 0003-2697 ,

    초록

    Abstract Readily accessible samples such as urine or blood are seemingly ideal for differentiating and stratifying patients; however, it has proven a daunting task to identify reliable biomarkers in such samples. Noncoding RNA holds great promise as a source of biomarkers distinguishing physiologic wellbeing or illness. Current methods to isolate and characterize RNA molecules in urine are limited. In this proof of concept study, we present a method to extract and identify small noncoding RNAs in urine. Initially, quantitative reverse transcription PCR was applied to confirm the presence of microRNAs in total RNA extracted from urine. Once the presence of micro RNA in urine was confirmed, we developed a method to scale up RNA extraction to provide adequate amounts of RNA for next generation sequence analysis. The method described in this study is applicable to detecting a broad range of small noncoding RNAs in urine; thus, they have wide applicability for health and disease analyses.

    원문보기

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    무료다운로드 유료다운로드

    회원님의 원문열람 권한에 따라 열람이 불가능 할 수 있으며 권한이 없는 경우 해당 사이트의 정책에 따라 회원가입 및 유료구매가 필요할 수 있습니다.이동하는 사이트에서의 모든 정보이용은 NDSL과 무관합니다.

    NDSL에서는 해당 원문을 복사서비스하고 있습니다. 아래의 원문복사신청 또는 장바구니담기를 통하여 원문복사서비스 이용이 가능합니다.

    이미지

    Fig. 1 이미지
  4. [해외논문]   Designing binding kinetic assay on the bio-layer interferometry (BLI) biosensor to characterize antibody-antigen interactions   SCI SCIE

    Kamat, Vishal (Corresponding author.) , Rafique, Ashique
    Analytical biochemistry v.536 ,pp. 16 - 31 , 2017 , 0003-2697 ,

    초록

    Abstract The Octet biosensors provide a high-throughput alternative to the well-established surface plasmon resonance (SPR) and SPR imaging (SPRi) biosensors to characterize antibody-antigen interactions. However, the utility of the Octet biosensors for accurate and reproducible measurement of binding rate constants of monoclonal antibodies (mAbs) is limited due to challenges such as analyte rebinding, and mass transport limitation (MTL). This study focuses on addressing these challenges and provides experimental conditions to reliably measure kinetics of mAb-antigen interactions. The mAb capture density of less than 0.6 nm was found to be optimal to measure a wide range of binding affinities on Octet HTX biosensor. The titration kinetic and single cycle kinetic assays performed on Octet HTX generated reproducible binding kinetic parameters and correlated with the values measured on Biacore 4000 and MASS-1. Kinetic assays performed on 0.1 nm density mAb surfaces significantly reduced MTL and enabled characterization of picomolar affinity mAbs. Finally, kinetic analysis performed on 150 antibodies to 10 antigens with molecular weights ranging from 21kD to 105kD showed concordance between Octet HTX, Biacore 4000 and MASS-1 (R 2 > 0.90). The data presented in this study suggest that under optimal experimental conditions, Octet biosensor is capable of generating kinetic values comparable to SPR/SPRi biosensors. Highlights High density antibody capture surface exhibited analyte rebinding and mass transport limitation (MTL). Kinetic assays performed on 0.1 nm density antibody surface significantly reduced MTL and enabled characterization of picomolar affinity interactions. A total of 165 fully human antibodies binding to 14 different antigens with molecular weight ranging from 14kD to 105kD were characterized on Octet HTX, Biacore 4000 and MASS-1 and the results revealed that measured binding kinetic values were comparable across different biosensors.

    원문보기

    원문보기
    무료다운로드 유료다운로드

    회원님의 원문열람 권한에 따라 열람이 불가능 할 수 있으며 권한이 없는 경우 해당 사이트의 정책에 따라 회원가입 및 유료구매가 필요할 수 있습니다.이동하는 사이트에서의 모든 정보이용은 NDSL과 무관합니다.

    NDSL에서는 해당 원문을 복사서비스하고 있습니다. 아래의 원문복사신청 또는 장바구니담기를 통하여 원문복사서비스 이용이 가능합니다.

    이미지

    Fig. 1 이미지
  5. [해외논문]   Real-time qRT-PCR assay for the detection of miRNAs using bi-directional extension sequences   SCI SCIE

    Kim, Kyung Jin (Heim Biotek, 35 Pangyo-ro 255 Beon-gil, Bundang-gu, Seongnam-si, Kyeonggi-do 13468, Republic of Korea ) , Kwak, Jiwon (Department of Pharmaceutical Engineering, Soonchunhyang University, 22 Soonchunhyang-ro, Shinchang-myeon, Asan-si, Chungcheongnam-do 31538, Republic of Korea ) , Lee, Jae-Hoon (Heim Biotek, 35 Pangyo-ro 255 Beon-gil, Bundang-gu, Seongnam-si, Kyeonggi-do 13468, Republic of Korea ) , Lee, Soo Suk (Department of Pharmaceutical Engineering, Soonchunhyang University, 22 Soonchunhyang-ro, Shinchang-myeon, Asan-si, Chungcheongnam-do 31538, Republic of Korea)
    Analytical biochemistry v.536 ,pp. 32 - 35 , 2017 , 0003-2697 ,

    초록

    Abstract Highly specific detection of miRNAs was performed using a novel bi-directional extension (BDE) assay. After reverse transcription, the cDNA was hybridized to a uniquely designed specific BDE sequence; this cDNA-BDE hybrid forms the PCR template. The PCR template was amplified in a SYBR Green-based quantitative real-time PCR. The miR-145 in human brain total RNA could be detected quantitatively in the range of seven orders of magnitude with high linearity and reproducibility. This innovative BDE assay has several performance advantages over the poly(A) tailing method that include lower C T values, clear gel electrophoresis images, and distinct nucleotide peaks in sequencing chromatograms.

    원문보기

    원문보기
    무료다운로드 유료다운로드

    회원님의 원문열람 권한에 따라 열람이 불가능 할 수 있으며 권한이 없는 경우 해당 사이트의 정책에 따라 회원가입 및 유료구매가 필요할 수 있습니다.이동하는 사이트에서의 모든 정보이용은 NDSL과 무관합니다.

    NDSL에서는 해당 원문을 복사서비스하고 있습니다. 아래의 원문복사신청 또는 장바구니담기를 통하여 원문복사서비스 이용이 가능합니다.

    이미지

    Fig. 1 이미지
  6. [해외논문]   DNA aptamer identification and characterization for E. coli O157 detection using cell based SELEX method   SCI SCIE

    Amraee, Masoum (Corresponding author.) , Oloomi, Mana , Yavari, Afsaneh , Bouzari, Saeid
    Analytical biochemistry v.536 ,pp. 36 - 44 , 2017 , 0003-2697 ,

    초록

    Abstract Escherichia coli (E. coli) O157:H7 is a foodborne pathogen that causes symptoms in humans. Its rapid identification should be considered to avoid toxic effects of the pathogen. In this study, systematic evolution of ligands by exponential enrichment using whole cells (Cell-SELEX) method was used for recognizing E. coli strain, O157 by single-stranded DNA library of aptamer. Nine rounds of cell-selex procedure were applied using O157, as a whole-cell target, with O42, K12, Top10, DH5α E. coli cells, Shigella flexneri and Salmonella typhi as counterparts. The specific interaction between selected DNA aptamers and targeted cell was assessed. After applying six rounds of SELEX for selection of DNA aptamers, the candidate sequences were obtained. Finally, specific aptamer was selected as an ideal aptamer for detection and capturing of E. coli O157. Dissociation constant of the selected aptamer were calculated (107.6 ± 67.8 pM). In addition, the secondary structure prediction and cross reactivity assays were performed. The isolated aptamer efficiency was confirmed and it was shown that the new DNA aptamer sequence has the ability to use for detection. This specific O157:H7 aptamer have the potential for application as a diagnostic ligand and could be used for detection of the related food borne diseases.

    원문보기

    원문보기
    무료다운로드 유료다운로드

    회원님의 원문열람 권한에 따라 열람이 불가능 할 수 있으며 권한이 없는 경우 해당 사이트의 정책에 따라 회원가입 및 유료구매가 필요할 수 있습니다.이동하는 사이트에서의 모든 정보이용은 NDSL과 무관합니다.

    NDSL에서는 해당 원문을 복사서비스하고 있습니다. 아래의 원문복사신청 또는 장바구니담기를 통하여 원문복사서비스 이용이 가능합니다.

    이미지

    Fig. 1 이미지
  7. [해외논문]   Enhanced post wash retention of combed DNA molecules by varying multiple combing parameters   SCI SCIE

    Yadav, Hemendra (Department of Botany, University of Rajasthan, Jaipur, India ) , Sharma, Pulkit (Bioinventree Pvt Ltd., SB 17 Bhawani Singh Road, Bapunagar, Jaipur 302015 India)
    Analytical biochemistry v.536 ,pp. 45 - 50 , 2017 , 0003-2697 ,

    초록

    Abstract Recent advances in genomics have created a need for efficient techniques for deciphering information hidden in various genomes. Single molecule analysis is one such technique to understand molecular processes at single molecule level. Fiber- FISH performed with the help of DNA combing can help us in understanding genetic rearrangements and changes in genome at single DNA molecule level. For performing Fiber-FISH we need high retention of combed DNA molecules post wash as Fiber-FISH requires profuse washing. We optimized combing process involving combing solution, method of DNA mounting on glass slides and coating of glass slides to enhance post-wash retention of DNA molecules. It was found that average number of DNA molecules observed post-wash per field of view was maximum with our optimized combing solution. APTES coated glass slides showed lesser retention than PEI surface but fluorescent intensity was higher in case of APTES coated surface. Capillary method used to mount DNA on glass slides also showed lesser retention but straight DNA molecules were observed as compared to force flow method.

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    원문보기
    무료다운로드 유료다운로드

    회원님의 원문열람 권한에 따라 열람이 불가능 할 수 있으며 권한이 없는 경우 해당 사이트의 정책에 따라 회원가입 및 유료구매가 필요할 수 있습니다.이동하는 사이트에서의 모든 정보이용은 NDSL과 무관합니다.

    NDSL에서는 해당 원문을 복사서비스하고 있습니다. 아래의 원문복사신청 또는 장바구니담기를 통하여 원문복사서비스 이용이 가능합니다.

    이미지

    Fig. 1 이미지
  8. [해외논문]   Graphene oxide layer decorated gold nanoparticles based immunosensor for the detection of prostate cancer risk factor   SCI SCIE

    Pal, Mintu (Biotechnology Group, Biological Sciences and Technology Division, CSIR-North East Institute of Science & Technology, Academy of Scientific and Innovative Research, Jorhat, 785006, Assam, India ) , Khan, Raju (Analytical Chemistry Group, Chemical Sciences and Technology Division, CSIR-North East Institute of Science & Technology, Academy of Scientific and Innovative Research, Jorhat, 785006, Assam, India)
    Analytical biochemistry v.536 ,pp. 51 - 58 , 2017 , 0003-2697 ,

    초록

    Abstract In this work, we report a novel electrochemical immunosensor based on gold nanoparticles deposited on the surface of graphene oxide layers and was used for immobilization of monoclonal anti-PSA antibody via EDC/NHS coupling method to detect prostate-specific antigen (PSA), a valuable biomarker for early detection of prostate cancer. To confirm the functionality of the antibody, we performed immunofluorescence staining using human prostate adenocarcinoma cells, LNCaP. Scanning Electron Microscopy (SEM), cyclic voltammetry, and other electrochemical techniques were used to characterize the resulting electrode surface. Unlike the previous research, this novel immunosensor functions very well with a low detection limit of 0.24 fgmL −1 at signal to noise ratio of 3; furthermore, it exhibits a significantly increased electron transfer and high sensitivity of 5.4 μA fgmL −1 with regression coefficient (R 2 = 0.99) toward PSA. The immunosensor was verified for selective and accurate detection of PSA in human serum with recovery of 97.67%. Overall, data suggested that our developed biosensor holds great promise as a useful alternative diagnostic tool for the detection of different cancer biomarkers, in particular PSA present in biological samples.

    원문보기

    원문보기
    무료다운로드 유료다운로드

    회원님의 원문열람 권한에 따라 열람이 불가능 할 수 있으며 권한이 없는 경우 해당 사이트의 정책에 따라 회원가입 및 유료구매가 필요할 수 있습니다.이동하는 사이트에서의 모든 정보이용은 NDSL과 무관합니다.

    NDSL에서는 해당 원문을 복사서비스하고 있습니다. 아래의 원문복사신청 또는 장바구니담기를 통하여 원문복사서비스 이용이 가능합니다.

    이미지

    Fig. 1 이미지
  9. [해외논문]   A miniaturized peptidyl-prolyl isomerase enzyme assay   SCI SCIE

    Vivoli, Mirella (College of Life and Environmental Sciences, University of Exeter, Exeter EX4 4QD, United Kingdom ) , Renou, Julien (Normandie Univ, UNIROUEN, CNRS, INSAREOUEN, COBRA, UMR 6014 & FR 3038, 1 rue Tesnière 76000 Rouen, France ) , Chevalier, Arnaud (Normandie Univ, UNIROUEN, CNRS, INSAREOUEN, COBRA, UMR 6014 & FR 3038, 1 rue Tesnière 76000 Rouen, France ) , Norville, Isobel H. (Defence Science and Technology Laboratory, Porton Down SP4 0JQ, United Kingdom ) , Diaz, Suraya (College of Life and Environmental Sciences, University of Exeter, Exeter EX4 4QD, United Kingdom ) , Juli, Christina (Institute of Pharmacy, University of Würzburg, Am Hubland, 970074 Würzburg, Germany ) , Atkins, Helen (Defence Science and Technology Laboratory, Porton Down SP4 0JQ, United Kingdom ) , Holzgrabe, Ulrike (Institute of Pharmacy, University of Würzburg, Am Hubland, 970074 Würzburg, Germany ) , Renard, Pierre-Yves (Normandie Univ, UNIROUEN, CNRS, INSAREOUEN, COBRA, UMR 6014 & FR 3038, 1 rue Tesnière 76000 Rouen, France ) , Sarkar-Tyson, Mitali (Defence Science and Technology Laboratory, Porton Down SP4 0JQ, United Kingdom ) , Harmer, Nicholas J. (College of Life and En)
    Analytical biochemistry v.536 ,pp. 59 - 68 , 2017 , 0003-2697 ,

    초록

    Abstract Prolyl-peptidyl isomerases (PPIases) are enzymes that are found in all living organisms. They form an essential part of the cellular protein folding homeostasis machinery. PPIases are associated with many important human diseases, e.g. cardiovascular disease, cancer and Alzheimer's. The development of novel PPIase inhibitors has been limited by the lack of a rapid, laboratory-based assay for these enzymes, as their substrates and products are challenging to distinguish. A well described continuous assay, coupled with the hydrolysis of a peptide by chymotrypsin is highly effective, but comparatively slow. To address this, we developed an improved version of the traditional assay using a temperature controlled plate reader. This assay allows semi-automated medium throughput assays in an academic laboratory for 84 samples per day. The assay shows lower errors, with an average Z′ of 0.72. We further developed the assay using a fluorogenic peptide-based FRET probe. This provides an extremely sensitive PPIase assay using substrate at 200 nM, which approaches single turnover conditions. The fluorescent probe achieves an excellent quenching efficiency of 98.6%, and initial experiments showed acceptable Z′ of 0.31 and 0.30 for cyclophilin A and hFKBP12 respectively. The assays provide an improved toolset for the quantitative, biochemical analysis of PPIases.

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    무료다운로드 유료다운로드

    회원님의 원문열람 권한에 따라 열람이 불가능 할 수 있으며 권한이 없는 경우 해당 사이트의 정책에 따라 회원가입 및 유료구매가 필요할 수 있습니다.이동하는 사이트에서의 모든 정보이용은 NDSL과 무관합니다.

    NDSL에서는 해당 원문을 복사서비스하고 있습니다. 아래의 원문복사신청 또는 장바구니담기를 통하여 원문복사서비스 이용이 가능합니다.

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  10. [해외논문]   Expression and purification of functional insulin and insulin-like growth factor 1 holoreceptors from mammalian cells   SCI SCIE

    Delle Bovi, Richard J. (Department of Physiology and Biophysics, Stony Brook University, NY, United States ) , Miller, W. Todd (Department of Physiology and Biophysics, Stony Brook University, NY, United States)
    Analytical biochemistry v.536 ,pp. 69 - 77 , 2017 , 0003-2697 ,

    초록

    Abstract The insulin receptor (IR) and insulin-like growth factor 1 receptor (IGF1R) are receptor tyrosine kinases (RTKs) involved in the regulation of many important cellular processes. The current proposed models of activation are derived from structural studies using soluble extracellular domains and cytoplasmic tyrosine kinase domains. Preparations of full length IR and IGF1R have been hampered by the need for unconventional affinity chromatography resins and/or harsh eluting conditions. Here, we present a purification protocol to obtain full-length, detergent solubilized IR and IGF1R at quantities suitable for biochemical and structural characterization. We screened a panel of 24 structurally diverse detergents for optimal ligand activation. The receptors purified in n-dodecyl-β-D-maltoside showed ligand-stimulated autophosphorylation and kinase activity, suggesting an intact transmembrane signaling mechanism. This convenient purification protocol can be used to produce high quantities of IR, IGF1R, or other RTKs, and can be adapted for other challenging membrane proteins.

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    원문보기
    무료다운로드 유료다운로드

    회원님의 원문열람 권한에 따라 열람이 불가능 할 수 있으며 권한이 없는 경우 해당 사이트의 정책에 따라 회원가입 및 유료구매가 필요할 수 있습니다.이동하는 사이트에서의 모든 정보이용은 NDSL과 무관합니다.

    NDSL에서는 해당 원문을 복사서비스하고 있습니다. 아래의 원문복사신청 또는 장바구니담기를 통하여 원문복사서비스 이용이 가능합니다.

    이미지

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