uses separation methods and separation principles
cell separation methods and applications
Ngày tải lên :
10/04/2014, 11:13
... molecular and cellular biology and genetics (Siebenkotten and co-workers) and the clinical isolation of stem and progenitor cells for cell therapy of cancer and other diseases (Hassan and co-workers) ... in cell biology, immunology, and molecular biology This book on Cell Separation Methods and Applications describes all of the important methods for the analytical and preparative isolation of ... centrifugal elutriation (Van Vlasselaer and coworkers) and density gradient separations (Figdor and colleagues) All of the more specific cell separation methods, based on monoclonal antibodies...
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Social Science Research - Principles, Methods, and Practices pptx
Ngày tải lên :
05/03/2014, 22:20
... education, and firm size, are easy to understand, others, such as creativity, prejudice, and organizational agility, may be more complex and abstruse, and still others such as trust, attitude, and learning, ... these effects are of a random (non-systematic) nature Two types of randomization are: (1) random selection, where a sample is selected randomly from a population, and (2) random assignment, where ... Quarterly and is frequently invited to present his research at universities and conferences worldwide Dr Bhattacherjee holds Ph.D and MBA degrees from the University of Houston, USA and M.S and B.S...
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electrochemistry principles, methods and applications 1993 - brett & brett
Ngày tải lên :
16/04/2014, 11:21
... Cataloging in Publication Data Brett, Christopher M A Electrochemistry: principles, methods, and applications/ Christopher M A Brett and Ana Maria Oliveira Brett Includes bibliographical references ... The sections are: Chapters 2-6 on the thermodynamics and kinetics of electrode reactions, Chapters 7-12 on experimental strategy and methods, and Chapters 13-17 on applications Also included are ... Secondly to our many colleagues and students who, over the years, with their comments and questions, have aided us in deepening our understanding of electrochemistry and seeing it with different...
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Glycoprotein Methods and Protocols - P1
Ngày tải lên :
08/10/2012, 10:25
... a UV monitor and collect fractions using a fraction collector and subject to the appropriate carbohydrate and antibody analyses 3.6 Analysis of Mucins Methods for the detection and analysis of ... antibodies and lectins or coating methods such as the glycan detection method and enzyme-linked immunosorbent assays (ELISA) All these techniques have advantages and disadvantages Solution methods ... remaining tissue From: Methods in Molecular Biology, Vol 125: Glycoprotein Methods and Protocols: The Mucins Edited by: A Corfield © Humana Press Inc., Totowa, NJ Davies and Carlstedt To isolate...
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Glycoprotein Methods and Protocols - P2
Ngày tải lên :
08/10/2012, 10:25
... mammary gland, defined by antibodies to cytokeratins J Cell Sci 75, 17–33 Kiernan, J A (1990) Histological and Histochemical Methods: Theory and Practice, Pergamon, Oxford, UK Andrew, S M and Jasani, ... M and Milton, J D (1986) Lectin Methods and Protocols, Humana, Totowa, NJ Liener, I E., Sharon, N., and Goldstein, I J (1998) The Lectins: Properties, Functions, and Applications in Biology and ... such as Culling’s Handbook of Histopathological and Histochemical Techniques (including museum techniques) (36) or Kiernan’s Histological and Histochemical Methods: Theory and Practice (45) provide...
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Glycoprotein Methods and Protocols - P3
Ngày tải lên :
08/10/2012, 10:25
... NJ Nagma, K., Thornton, D J., Khan, N., and Sheehan, J K (2000) Separation and identification of mucins and their glycoforms, in Glycoprotein Methods and Protocols: The Mucins (Corfield, T., ... greater sample volume, and superior washing) For quantitation and comparison across blots, a standard in the same buffer as samples should be titrated for use as a standard curve, and samples should ... core protein specificity), and lower sensitivity than slot-blot and immunoassay methods Chap- Detection and Quantitation of Mucins 47 ter (2) discusses these assays and they are mentioned here...
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Glycoprotein Methods and Protocols - P4
Ngày tải lên :
08/10/2012, 10:25
... use of fixatives and organic solvents together with conventional paraffin embedding causes denaturation, dehydration, and loss of the surface mucous gel layer (9–11) Similarly, standard preparation ... and held together by bolts tightened by hand Millipore filter paper to act as a backing for the mucosa during sectioning and subsequent manipulation of the section 2.2 Histological Staining and ... (w/v) poly-L-lysine for and dried at room temperature Methods 3.1 Observation and Measurement of Mucous Thickness on Unfixed Mucosal Sections (see Notes 1–4) This is a rapid and simple method for...
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Glycoprotein Methods and Protocols - P5
Ngày tải lên :
08/10/2012, 10:25
... precursor as arbitrary units (au)/milliliter of homogenate Identify and quantify the mature mucin band in the homogenate and medium after separation on reducing 4% SDS-PAGE using the PhosphorImager (see ... PhosphorImager The identification of mucin precursors and mature mucins using polypeptide-specific antisera and SDS-PAGE is elaborated in Chapters 20 and 21, and in several references (1,4–11) The precursor ... Strous, G J and Dekker, J (1992) Mucin-type glycoproteins Crit Rev Biochem Mol Biol 27, 57–92 Van Klinken, B J W., Dekker, J., Büller, H A., and Einerhand, A W C (1995) Mucin gene structure and expression...
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Glycoprotein Methods and Protocols - P6
Ngày tải lên :
08/10/2012, 10:25
... continuum in the electro- Separation and Identification of Mucin 81 Figure Separation of reduced mucin subunits: (A) Mucins were isolated from saliva, reduced and alkylated, and chromatographed on ... a small gel, use 1.6 g of agarose and 160 mL of buffer, and for a large gel, use 2.8 g of agarose and 280 mL of buffer Leave to cool before pouring (hand hot) and insert well-forming comb (see ... are 1.5 mm thick and cm wide There is a compromise between band broadness and amount of sample to be loaded Larger amounts of sample tend to yield poorer quality data; i.e., the bands become more...
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Glycoprotein Methods and Protocols - P7
Ngày tải lên :
08/10/2012, 10:25
... Kadler, K E., and Chapman J A (1985) Mica sandwhich technique for preparing macromolecules for rotary shadowing J Ultrastruct Res 91, 66–76 14 Kawahara K and Tanford C (1966) Viscosity and density ... temperature and mechanical instabilities in centrifuges We first demonstrated the value of rate-zonal centrifugation for the characterization of cervical mucin heterogeneity and polydispersity (5) and ... (Fullerton, CA) SW 40Ti swing-out rotor, and finally unloading and analysis of the tube contents Measure out 18 mL of M GuHCl into one chamber of the gradient maker and an equal volume of M GuHCl into...
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Glycoprotein Methods and Protocols - P8
Ngày tải lên :
08/10/2012, 10:25
... at least for pump and column equilibration with the initial composition of the mobile phase Figures and show typical chromatograms of the separation of amino acid standards and hydrolyzed bovine ... 114 Yan and Packer chromatography, and leads to a significantly lower recovery of all the amino acids, especially serine and threonine Strong acid, high temperature, and short time acid ... samples and Dr Andrew Gooley and Prof Keith Williams for their support References Yan, J X., Wilkins, M R., Ou, K., Gooley, A A., Williams, K L., Sanchez, J.-C., Golaz, O., Pasquali, C., and Hochstrasser,...
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Glycoprotein Methods and Protocols - P9
Ngày tải lên :
08/10/2012, 10:25
... Notes and 8) and the length of sequence obtained is dependent on the number and distribution of proline residues Proline cleaves with reduced efficiency in Edman sequencing (Fig 1, vessel 1), and ... I., Kirk, M., Barnes, S., Lowary, T L., and Hart, G W (1996) Selective detection and site-analysis of O-GlcNAc-modified glycopeptides by β-elimination and tandem electrospray mass spectrometry Anal ... acids modified by monosaccharides and larger oligosaccharides (degree of polymerization [DP]=19) Elsewhere we have published alternative HPLC methods for the separation of PTH-glycosylated amino...
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Glycoprotein Methods and Protocols - P10
Ngày tải lên :
08/10/2012, 10:25
... tandem repeat domain and overlapping each other by six amino acids (16), and the other used octamers overlapping each other by seven amino acids (17) Twelve antibodies were analyzed in total and ... information on antibody specificity and permits identification of significant features of an epitope that may contribute to antibody recognition and binding (see Notes 10 and 11) 1.3 Purification of ... must be pure and free from amines Several methods may be used to purify DMF (see Note 16) Piperidine 20% v/v: used for Fmoc deprotection Piperidine should be redistilled before use and made up...
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Glycoprotein Methods and Protocols - P11
Ngày tải lên :
08/10/2012, 10:25
... lysates and media Nonspecific bands may be present and can be eliminated from consideration using the pSVL vector control lanes for comparison (Fig 2A,B, lanes and 3) The sizes of specific bands ... polymerase buffer, 2.5 mM MgCl2, 400 µM dNTPs, 200 ng of primers and or and 4, ng of template DNA (with only primers and 4), and U of Taq polymerase Perform a PCR program in a suitable thermal ... polymerase, and incubate the reaction at 72˚C to allow extension of the linked templates Add µg each of primers and 6, and continue the PCR with 30 cycles at 94˚C for 30 s, 50˚C for and 72˚C for...
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Glycoprotein Methods and Protocols - P12
Ngày tải lên :
08/10/2012, 10:25
... temperature from 150 to 230°C and 3°C/min and then 230 to 250°C at 5°C/min Use injector and detector temperatures of 240 and 250°C, respectively Use an injection volume of µL 3.3 Separation of Monosaccharides ... 164 Michalski and Capon Fig HPLC separation of sialic acid quinoxalinones obtained after mild acid hydrolysis of BSM Ac, acetyl; Lt, lactyl; Gc, glycolyl Methods 3.1 Release and Identification ... temperature from 120 to 240°C at 2°C/min Use injector and detector temperatures of 240 and 250°C, respectively 168 168 Michalski and Capon Fig GLC separation of trimethylsilylated deriviatives of...
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Glycoprotein Methods and Protocols - P13
Ngày tải lên :
08/10/2012, 10:25
... (100 × cm) in a water jacket at 55°C and HPLC pump with refractive index and fluorescence detectors GlycoSepH™ and GlycoSepC™ HPLC column (Oxford GlycoSciences) Methods 3.1 Alkaline Borohydride Degradation ... 6.0, and load the sample carefully and directly to the center of the column, drop wise (see Note 3) Centrifuge for at 1100g and collect the excluded O-Glycanase O-Linked Chain Release and Fractionation ... (3 × 0.5 cm column) and elute with three column vol of HPLC-grade water Collect the effluent and eluent and then pool (2.5-mL volume) Load onto a PD10-Sephadex GM25 column and elute with HPLC-grade...
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Glycoprotein Methods and Protocols - P14
Ngày tải lên :
08/10/2012, 10:25
... 2-aminopyridine, and separation and characterization of the derivatives by ESI-MS/MS First, the mild periodate oxidation (10 nmol of oligosaccharide-alditols disssolved in an imidazole buffer, and reacted ... appearence of novel methods of ionization such as fast atom bombardment ionization (FAB), electrospray ionization (ESI), and matrixassisted laser desorption ionization (MALDI) Methods 2.1 Nuclear ... method was developed by Vliegenthart and colleagues during the 1970s and essentially applied to the structure determination of N-glycans of the Nacetyllactosamine and oligomannoside type (16) More...
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Glycoprotein Methods and Protocols - P15
Ngày tải lên :
08/10/2012, 10:25
... hygroscopic and must be thoroughly dried before weighing and making up as standards Standards of mM each of 2-, 3-, and 6-sulfo-Nacetylglucosamine in water may be aliquoted and stored at –20°C, and used ... chloride (Cl–), sulfate (HSO4–), and phosphate (H2PO3–) ions (see also Note 11) Chromatogram displays 10 nmol each of phosphate and sulfate, and trace amounts of chloride and acetate Note the difference ... 212 Harrison and Packer Materials 2.1 Solvents and Reagents Water, carbonate-free (see Note 1), and sparged with a constant flow of argon 0.1 M NaOH, made up with decarbonated water, and similarly...
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Glycoprotein Methods and Protocols - P16
Ngày tải lên :
08/10/2012, 10:25
... Subheading 2., item 5e at 37°C and 5% CO2 Caco-2 and A431 cells are passaged (trypsinised) and split 1:4 and the medium is changed every to d LS174T cells are split 1:2 and the medium changed daily ... only MUC and MUC2 at early passage, but at later passage and in later premalignant and malignant stages, they show de novo expression of MUC5AC, MUC5B, and MUC6 (Corfield, Myerscough, and Paraskeva, ... clumps of cells and replate under standard culture conditions Reattachment of cells may take several days, and during medium changing, any floating clumps of cells must be centrifuged and replated...
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Glycoprotein Methods and Protocols - P17
Ngày tải lên :
08/10/2012, 10:25
... Howard, M., Devine, P L., and Sheehan, J K (1995) Methods for separation and deglycosylation of mucin subunits Anal Biochem 227, 162–167 Corfield, A P., Clamp, J R., Casey, A D., and Paraskeva, C (1990) ... After incubation, remove the medium from the central well and wash the tissue and dish with mL of PBS Pool the medium and washings, and dialyze against three changes of L of M guanidine hydrochloride ... Electrophoresis and Vacuum Blotting (see Notes 16 and 17) Mix samples containing 10–500 mg mucin or >10,000cpm with 50 mL sample buffer and load onto horizontal 0.8–1% agarose gels Run Rainbow markers and...
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