carbon membrane derived from interfacial charged grafted double polymer layers for gas separation
Macro to micro porous ceramic and carbon media philosophy of design and fabrication
... 12, Carbon membrane derived from interfacial charged- grafted double polymer layers for gas separation , we developed a new fabrication concept for the carbon membrane using conducting polymer ... 267 11.5 Conclusions 270 Chapter 12 : Carbon Membrane derived from Interfacial Charged- grafted Double Polymer Layers for Gas Separation 271 12.1 Introduction ... 8-13: Membrane performance for 10wt% membrane at atm: (a) Permeation flux for pristine membrane (b) Rejection for pristine membrane (c) Permeation flux for CNT membrane (d) Rejection for CNT membrane...
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Safe carrying methods
... accidentally removing the mucous from fish Gloves can also protect you from pricks and cuts from broken branches or rubbish that find their way into and around the pond, or from contracting diseases ... substance or work practice) Separate the hazard from people with the use of screens, safety barriers, or find another place away from people Use safeguards for tools, equipment, machinery or install ... in the way of fines for not following workplace health and safety legislation These principles are controlled by laws that differ from state to state In Western Australia, for example, these principles...
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Glycoprotein methods protocols - biotechnology 048-9-001-013.pdf
... Isolation of Large Gel-Forming Mucins Isolation of Large Gel-Forming Mucins Julia R Davies and Ingemar Carlstedt Introduction The large gel-forming mucins, which form the major macromolecular ... large, gel-forming glycoproteins and smaller, monomeric ones The gel-forming mucins (Mr = 10–30 million Dalton) are oligomers formed by subunits (monomers) joined via disulfide bonds (for a review ... epithelium For example, material from the surface epithelium may be enriched by gently scraping the surface mucosa, thereby allowing gland material to be obtained from the remaining tissue From: Methods...
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Glycoprotein methods protocols - biotechnology 048-9-027-044.pdf
... distilled water Oxidize for 5-15 in 1% periodic acid Wash in running tap water for and then rinse in distilled water Treat sections with Schiff’s reagent for 10–30 Wash for 10 in running tap water ... changes for each Block endogenous peroxidase activity using 1.0% H2O2 and 0.1% NaN3 in PBS for 10 Wash in three changes of PBS for each Incubate sections in 4% commercial skim milk powder in PBS for ... 1:300–1:500 for 30 Wash in three changes of PBS for each Incubate sections with streptavidin-horseradish peroxidase (HRP) conjugate (Jackson) diluted 1:250–1:400 for 30 Wash in three changes of PBS for...
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Glycoprotein methods protocols - biotechnology 048-9-045-055.pdf
... feature that can be an advantage for one application could be a disadvantage for another application For example, use of specific peptide-reactive antibodies for known mucin core proteins may ... DETECTION For enzyme assays, the choice of substrate and buffer depends on the enzyme: ABTS, OPD, or TMB for HRP; PNPP for alkaline phosphatase (AP) Incubate at room temperature or 37°C for 20–60 ... times for each in sodium metabisulfite and dry the membrane in a warm airstream 3.2.3 Detection of Mucins on Membranes Using Antibodies or Lectins (see Notes 10,12 and 13) 3.2.3.1 BLOCKING Membranes...
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Glycoprotein methods protocols - biotechnology 048-9-057-064.pdf
... negatively charged nucleic acids and cell surface components, will bind the dye, and it has yet to be shown that the dye penetrates the mucous layer uniformly Results from this method are therefore ... other in vitro methods for measuring mucous thickness, for studying mucous secretion in vivo, and for studying the secretion of acid from the gastric glands through the covering mucous layer (20) ... range Therefore, results are best treated for statistical significance by methods that compare nonparametric distributions such as the Mann-Whitney U-test Values for thickness should therefore be...
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Glycoprotein methods protocols - biotechnology 048-9-065-073.pdf
... mucin of interest for 90 b Wash two times for 15 each c Incubate with 0.5 µCi of (185 kBq) 125I-labeled protein A for 60 d Wash twice for each in Blotto, and then wash once for in phosphatebuffered ... aprotinin 1% (v/v) form commercial solution, mM phenylmethylsulfonylfluoride (PMSF), 10 µg/mL leupeptin All chemicals are from Sigma PMSF is unstable in water; add just before use, from 100 mM stock ... and air-dry Immerse the filter paper in ice-cold 5% TCA for at least 10 Transfer paper to 5% TCA at 100°C for 10 Wash the paper two times for each in 5% TCA at room temperature Rinse once in 50%...
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Glycoprotein methods protocols - biotechnology 048-9-075-085.pdf
... is typical of that observed for the reduced subunits prepared from the gel-forming mucins isolated from other epithelial secretions (respiratory and cervical) and from intestinal cell lines (HT-29 ... differently charged forms of this mucin The continuum in the electro- Separation and Identification of Mucin 81 Figure Separation of reduced mucin subunits: (A) Mucins were isolated from saliva, ... 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 Note 2) Leave to set for at least h prior...
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Glycoprotein methods protocols - biotechnology 048-9-087-096.pdf
... 2.230 2.400 values for ρ and ηrel are from ref 14 Linear gradient makers, suitable for making gradients of different volumes, can be purchased from a variety of manufacturers For the × 12 mL gradients ... Gel-Forming Mucins 95 For the Beckman SW 40Ti rotor, we typically centrifuge for 2.5 h to disperse intact mucins across the gradient and 6–8 h for the reduced subunits This is not usually performed ... microscopy of biopolymers Biopolymers 9, 373–379 96 Sheehan and Thornton 12 Koller, T., Harford, A G., Lee, Y K., and Beer, M (1969) New methods for the preparation of nucleic acid molecules for electron...
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Glycoprotein methods protocols - biotechnology 048-9-111-119.pdf
... vessel in a 105°C oven for 24 h Remove the vessel from the oven and open the vacuum tap immediately within a fume hood (see Note 4) Place the vials into a vacuum centrifuge for 10 to evaporate excess ... total amino acids x 100% (see Table for an example) (see Note 7) Amino Acid Analysis of Mucins 117 Fig Typical chromatogram of separation of amino acids from bovine conjunctival mucin hydrolysate ... their acid form: aspartic acid and glutamic acid, respectively) The mucin hydrolysate can be analyzed for its amino acid composition by any other chemical derivatization procedure and HPLC separation, ...
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Glycoprotein methods protocols - biotechnology 048-9-121-128.pdf
... water 2.2 Reagents for Adsorption onto Hyperbond Hyperbond membranes were from Beckman 2.3 Protein Coupling to Arylamine Membranes (see Note 1) Reagents are available in kit form from Beckman and ... monosaccharides and larger oligosaccharides (degree of polymerization [DP]=19) Elsewhere we have published alternative HPLC methods for the separation of PTH-glycosylated amino acids on a modified ... These methods were developed for two reasons: (1) to provide a distinct elution window for the PTH-glycosylated amino acids, and (2) to be of low enough salt concentration for the direct infusion...
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Glycoprotein methods protocols - biotechnology 048-9-129-141.pdf
... piperidine for 20 The piperidine solution should cover the gears The pins are then washed as follows: a DMF to cover gears for b MeOH (complete submersion) for c MeOH to cover gears for (three ... improved purification performance For instance, it has been found that the C595 mAb has higher affinity for a synthetic MUC1 glycopeptide compared with the naked peptide This information has been used ... chromatography text for further details NaSCN gradient elution provides a simple method for determining the relative affinities of a series of antibodies for a matrix or a single antibody for different...
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Glycoprotein methods protocols - biotechnology 048-9-143-155.pdf
... performed by the Biotechnology Service Centre, The Hospital for Sick Children, Toronto, Canada References Forstner, J F and Forstner, G G (1994) Gastrointestinal mucus, in Physiology of the Gastrointestinal ... results Wash the membrane as described in step Incubate the membrane for h in secondary incubation solution Wash the membrane as in step After a 3-min equilibration, develop the membrane in reaction ... in a suitable thermal cycler at 94˚C for min, followed by 30 cycles at 94˚C for min, 60˚C for min, and 72˚C for min, ending with an extension period of 72˚C for Couple the signal peptide and RMuc2...
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Glycoprotein methods protocols - biotechnology 048-9-159-180.pdf
... Solvents for HPAEC separation of sialic acids (see Subheading 3.3.3.3.) a Eluent 1: Deionized water b Eluent 2: mM NaOAc c Eluent 3: mM acetic acid (glacial, HPLC grade; Merck) 16 Solvents for separation ... 173 Table Elution Gradient for the Separation of Neutral Monosaccharides and Hexosamines Time (min) Eluent (%) Eluent (%) 94 94 6 30 Table Elution Gradient for the Separation of Neutral Monosaccharides, ... Table Elution Gradient for the Separation of Unreduced and Reduced Monosaccharides Time (min) 22 25 Eluent (%) 60 60 45 45 Eluent (%) 40 40 55 55 Table Elution Gradient for the Separation of GalNAc-ol...
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Glycoprotein methods protocols - biotechnology 048-9-181-190.pdf
... information More information about these techniques can be obtained from refs 18–20 The use of electrospray ionization MS coupled with collision-induced dissociation MS is another possibility for ... Oxford, GlycoSciences) 2.6 2-AB Fluorescence Labeling and Size Exclusion Chromatography (SEC) Signal labeling kit (K404,Oxford GlycoSciences) Dowex AG50 X 12 (H+ form) Dowex AG1 X (acetate form) ... acetonitrile for b 62.5% acetonitrile over the next 25 c 60% over the next 30 d Back to 75% at 60 for the total run 3.6.2 Porous Graphitized Carbon Chromatography Wash the column with water for 30...
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Glycoprotein methods protocols - biotechnology 048-9-191-209.pdf
... resulted from the connectivities 1IV/3V, 1V/4IV, 1IV/3III, 1III/4II, and 1II/3I Fig (continued) successively assigned The shape of the correlation peaks (triplet for H-2, pseudo-doublet for H-3, ... H-2, pseudo-doublet for H-3, pseudo-singlet for H-4) allows the estimatation of the coupling constant: L (large) for J > Hz; S (small) for J < Hz For the monosaccharide unit III, we observe J1,2 ... potential, and the solution emerges from the capillary as an aerosol of charged droplets Solvent evaporation through a nitrogen flux results in the formation of charged sample molecules, and the...
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Glycoprotein methods protocols - biotechnology 048-9-211-216.pdf
... nmoles of sulfate from 50 mg of mucin glycopeptides was established (see Note 10) Notes Carbonate ions (CO32-) elute at a retention time close to that of sulfate It is necessary to decarbonate water ... and B from human MUC2 The chromatogram shown corresponds to half of an original hydrolysis amount of 50 µg Our system for sulfate analysis uses the same chromatographic system as that for sugar ... amount of protein required for analysis will often be quite small For example, 10 pmol of a highly sulfated mucin (>100 mol sulfate/mol of protein) would be adequate for a confident analysis,...
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Glycoprotein methods protocols - biotechnology 048-9-219-226.pdf
... control for DNA laddering A mucosal biopsy is taken by endoscopy from a human individual with an otherwise intact organ A tissue explant is obtained from surgically resected human or animal tissue For ... at 300g for Roughly put cm3 in 20–40 mL of solution Rotate at 37°C overnight (12–16 h) Mix the suspension by using a Pasteur pipet to improve the separation of the epithelial elements from the ... conditions for adenoma cell lines as previously described for primary cultures Carry out passage of adenoma cells as clumps of cells using sufficient dispase just to cover the cells, and incubate for...
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Glycoprotein methods protocols - biotechnology 048-9-227-237.pdf
... 12,000g for at 4°C, and separate the supernatant soluble fraction from the membrane pellet Resuspend the membrane pellet in mL of M guanidine hydrochloride PBS/inhibitor cocktail 3.3 Separation ... identification of a lower risk group for severe disease and cancer Gut 36, 696–702 Forstner, J F and Forstner, G G (1994) Gastrointestinal mucus, in Physiology of the Gastrointestinal Tract (Johnson, ... to obtain a mucin-enriched fraction from crude culture samples for comparative studies (1,2) It is also a starting point for preparation of native mucins for further purification and analysis...
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