048 assimilation

048 assimilation

048   assimilation
... DE-ALVEOLAR ASSIMILATION 4 Prof Francisco Zabala - 2016 Phonemic changes 2.2.1 Bilabial assimilation The alveolar s+ c+ m may become bilabial before a bilabial sound Notice, however, that the assimilation ... (1992) 5 Prof Francisco Zabala - 2016 2.2.2 Velar assimilation The alveolar s+ c+ m may become velar before a velar sound Notice, however, that the assimilation of s into j is not as frequent as ... !adHj?m = !adHjMÈ TYPES OF CHANGE 2.1 Voicing Unlike many other languages, English does not frequently favour assimilation of voice In Spanish, on the contrary, voiceless sounds tend to become voiced in the...
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Glycoprotein methods protocols - biotechnology 048-9-001-013.pdf

Glycoprotein methods protocols - biotechnology 048-9-001-013.pdf
... different methods are available: solution assays such as colorimetric assays for hexose and sialic acid; membrane-based methods such as slot-blotting and staining with periodic acid-Schiff reagent; ... correct volume of 10 Davies and Carlstedt ice-cold dry propan-1-ol to give a 100 mM solution DFP is unstable in water but can be stored at –20°C in propan-1-ol After dilution, the vial as well as ... putative cell membrane-associated mucin Biochem J 338, 325–333 Desseyn, J.-L., Guyonet-Dupérat, V., Porchet, N., Aubert, J.-P., and Laine, A (1997) Human mucin gene MUC5B, the 10.7-kb large central...
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Glycoprotein methods protocols - biotechnology 048-9-027-044.pdf

Glycoprotein methods protocols - biotechnology 048-9-027-044.pdf
... to T-antigen (β-d-Gal 1-3 GalNAc), but also to structures found within the backbone of oligosaccharides (β-d-Gal 1-3 / 4GlcNAc) (19) Demonstration of PNA binding is not necessarily evidence of T-antigen ... Blue 8GX in 0.1 N HCl (pH 1.0) HID: Dissolve 120 mg of N,N-dimethyl-m-phenylenediamine dihydrochloride (Sigma) and 20 mg of N,N-dimethyl-p-phenylenediamine dihydrochloride (Sigma) in 50 mL distilled ... are, however, several other alternatives, such as 3-amino9-ethylcarbazole (AEC), for use with horseradish peroxidase (HRP); 5-bromo-4-chloro3-indolyl phosphate/nitro blue tetrazolium (BCIP/NBT);...
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Glycoprotein methods protocols - biotechnology 048-9-045-055.pdf

Glycoprotein methods protocols - biotechnology 048-9-045-055.pdf
... nonionic detergents: 0.05% (v/v) Nonidet P-40 or Tween-20 Enzyme substrates: 2,2'-azino-bis(3-ethylbenzathiazaline 6-sulfonic acid) (ABTS) (1 mg/mL, A405nm), O-phenyldiamine (OPD) (1 mg/mL, A492nm), ... chromogens that leave a colored precipitate on the blot (e.g., 3,3'-diaminobenzidine, 4-chloronapthol mix for HRP [14]; 5-bromo-4-chloro-3'-indolyphosphate toluidine salt, nitro blue tetrazolium chloride ... Histologically-based methods for detection of mucin, in Glycoprotein Methods and Protocols: The Mucins (Corfield, T., ed.), Humana, Totowa, NJ Davies, R., Carlstedt, I (2000) Isolation of large gel-forming...
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Glycoprotein methods protocols - biotechnology 048-9-057-064.pdf

Glycoprotein methods protocols - biotechnology 048-9-057-064.pdf
... 250 mg of phenol in 70 mL of glycerol Poly-L-lysine-coated slides: slides are immersed in 0.01% (w/v) poly-L-lysine for and dried at room temperature Methods 3.1 Observation and Measurement of ... A., Allen, A., Morris, E., and Ross-Murphy S B (1988) Mucus glycoprotein gels: role of glycoprotein polymeric structure and carbohydrate side-chains in gel-formation Carbohydrate Res 178, 93–110 ... prostaglandin, N-acetyl-cysteine and other agents Clin Sci 63, 187–195 Bell, A E., Allen, A., Morris, E R., and Ross-Murphy, S B (1984) Functional interactions of gastric mucus glycoprotein Int...
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Glycoprotein methods protocols - biotechnology 048-9-065-073.pdf

Glycoprotein methods protocols - biotechnology 048-9-065-073.pdf
... as [35S]sulfate-labeled, TCA-precipitable proteins (cpm/mL) (see Note 10) g Mature [35S]sulfate-labeled, mucin band on 4% SDS-PAGE (au/mL) h Total concentration of mucin by dot-blotting (au/mL) ... before use, from 100 mM stock solution in 2-propanol 21 Blotto (wash buffer for Western-type dot-blots): 50 mM Tris (pH 7.8), mM CaCl2, 0.05% (v/v) Nonidet P-40 (BDH), 0.01% antifoam A (Sigma), 5% ... radiolabel into (glyco-)proteins by TCA precipitation Spot µL of the homogenate on a pencil-marked location on 3MM Whatman filter paper and air-dry Immerse the filter paper in ice-cold 5% TCA for...
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Glycoprotein methods protocols - biotechnology 048-9-075-085.pdf

Glycoprotein methods protocols - biotechnology 048-9-075-085.pdf
... advisable We analyze aliquots of the peaks by using matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS), and purify peptides to homogeneity by rechromatography ... effectiveness of the separation is monitored with a periodic acid-Schiff (PAS) assay and with a variety of lectins and mucin- or carbohydrate-specific antibodies (for a discussion of the relative merits ... electrophoresis apparatus, e.g., Bio-Rad DNA subcell Model 96 (25 × 15 cm gels) and the Bio-Rad subcell (15 × 15 cm gels) Agarose (ultrapure) Electrophoresis buffer: 40 mM Tris-acetate, mM EDTA, pH 8.0,...
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Glycoprotein methods protocols - biotechnology 048-9-087-096.pdf

Glycoprotein methods protocols - biotechnology 048-9-087-096.pdf
... alternative, however, is rate-zonal centrifugation, which separates molecules not on hydrodynamic volume alone but on their mass-to-volume ratio as well The basis of rate-zonal centrifugation for ... Distribution of Gel-Forming Mucins 89 Fig Schematic diagram of gradient-forming apparatus We next describe an experiment that involves the simultaneous preparation of × 12 mL (in 13-mL tubes) 6–8 ... PREPARATION OF CARBON-COATED GRIDS Prepare a thin carbon film (2–5 nm) by the indirect evaporation of carbon onto 2-cm2 blocks of freshly cleaved mica (Fig 3) Leave the mica in a water-saturated environment...
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Glycoprotein methods protocols - biotechnology 048-9-111-119.pdf

Glycoprotein methods protocols - biotechnology 048-9-111-119.pdf
... of mucin glycoproteins The hydrolysate of mucin is then derivatized with Fmoc on the α-amino groups under alkaline conditions The derivatized amino acids are separated by a C18 reversed-phase column ... continuous flow of helium Hypersil C18 reversed-phase column: 150 × 4.6 mm inner diameter µm (Keystone, Bellefonte, PA) In-line filter (2 µm) (Upchurch, cat no 10 0-1 0) 2.2 Chemicals 2.2.1 Hydrolysis and ... Wilkins, M R., Ou, K., Gooley, A A., Williams, K L., Sanchez, J.-C., Golaz, O., Pasquali, C., and Hochstrasser, D.F (1996) Large-scale amino-acid analysis for proteome studies J Chromatogr 736, 291–302...
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Glycoprotein methods protocols - biotechnology 048-9-121-128.pdf

Glycoprotein methods protocols - biotechnology 048-9-121-128.pdf
... degradation: the identification of Xaa-Pro-Xaa-Xaa as a motif for Thr-O-glycosylation Biochem Biophys Res Commun 178, 1194–1201 Laursen, R A (1971) Solid-phase Edman degradation: an automated ... (C) The glycosylated amino acid window with elution profiles of PTH-threonine-GalNAc-Gal (black arrowheads) and PTH-threonine-GalNAc (gray arrowheads) from the Applied Biosystems Procise Standard ... arylamine-coated membranes are stable at –20°C; the N-ethyl-N'-dimethylaminopropylcarbodiimide (EDC reagent is stable at 4°C in a dry environment, and the buffer is stable at 4°C Arylamine-derivatized...
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Glycoprotein methods protocols - biotechnology 048-9-129-141.pdf

Glycoprotein methods protocols - biotechnology 048-9-129-141.pdf
... protecting group t-Butyl ether t-Butyl ester t-Butoxycarbonyl 2,2,5,7,8-Pentamethylchroman-6-sulfonyl Trityl Amino acid S, T, Y D, E K, H, W R C Materials 2.1 Preparation of BSA-Peptide Conjugates ... 16.8 g of citric acid monohydrate made up to L with distilled water, pH 4.0 2,2'-azino-bis[3-ethylbenz-thiazoline-6-sulfonic acid] (ABTS) substrate solution (Sigma): 0.5 mg/mL in citrate phosphate ... B., De Matteis, C I., Stadie, T and Hanisch, F.-G (1996) Effect of glycosylation of a synthetic MUC1 mucin-core-related peptide on recognition by anti-mucin antibodies Cancer Lett 100, 11–15 16...
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Glycoprotein methods protocols - biotechnology 048-9-143-155.pdf

Glycoprotein methods protocols - biotechnology 048-9-143-155.pdf
... sterilized 20 mM benzyl 2-acetamido-2-deoxy-α-D-galactopyranoside (benzyl-α-GalNAc) (Sigma), filter sterilized Peptide-N 4-( acetyl-ß-glycosaminyl) asparagine amidase (N-glycosidase F, EC 3.5.1.52) ... chloride and 1.65 mg of 5-bromo-4chloro-3-indolyl-phosphate), and stop solution (10 mM Tris-HCl, mM EDTA, pH 8.0) Nitro blue tetrazolium (NBT) and 5-bromo-4-chloro-3-indolyl-phosphate (BCIP) are ... G (1996) Benzyl-N-acetyl-α-D-galactosaminide inhibits the sialylation and the secretion of mucins by a mucin secreting HT-29 cell subpopulation Glycoconj J 13, 717–726 25 Perez-Vilar, J., Eckhardt,...
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Glycoprotein methods protocols - biotechnology 048-9-159-180.pdf

Glycoprotein methods protocols - biotechnology 048-9-159-180.pdf
... monosaccharides by reversed-phase HPLC Ultrasphere ODS (4.6 × 250 mm) 1, PA-Gal; 2, PA-Glc; 3, PA-Man; 4, PA-Rib; 5, PAFuc; 6, PA-Rham; 7, PA-ManNAc; 8, PA-deoxy-Rib; 9, PA-GlcNAz; 10, PA-GalNAc; 11, PANeuAc ... Oligosaccharide-alditols released by Type of reagents Detection Sensitivity HPLC Benzoyl p-Bromo-benzoyl; naphtoyl-benzoyl Dns-hydrazide DABS hydrazide Fmoc hydrazide Dansyl-hydrazide 1-phenyl-3-methyl-5-pyrazolone ... complexes of N-2-pyridyl-glycamines Anal Biochem 176, 72–77 21 Chiesa, C and Horvath, C (1993) Capillary zone electrophoresis of malto-oligosaccharides derivatized with 8-amino-naphtalene-1,3,6-trisulfonic...
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Glycoprotein methods protocols - biotechnology 048-9-181-190.pdf

Glycoprotein methods protocols - biotechnology 048-9-181-190.pdf
... normal- or reversed-phase high-performance liquid chromatography (HPLC) (8) or anion-exchange chromatography, e.g., high-performance anion-exchange chromatography (HPAEC) on two CarboPac PA-100 ... Dowex 50X-12 H+-form resin Sephadex GM25 2.4 Hydrazinolysis 0.5-mL screw-capped V-bottomed Reacti-vials™ (Pierce and Warriner, Chester, UK) Anhydrous hydrazine (Pierce and Warriner) Whatman CF-11 ... butan-2,3-diol (two times the molar excess over periodate) for 40 at 0°C in the dark 3.3 Release of Core Disaccharide (Galβ 1-3 GalNAcα–) from Mucin Using Endo-α-N-Acetylgalactosaminidase (O-Glycanase)...
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Glycoprotein methods protocols - biotechnology 048-9-191-209.pdf

Glycoprotein methods protocols - biotechnology 048-9-191-209.pdf
... GalNAc-ol H-2, H-4, H-5 and H-6' Analysis of Mucin-Type O-Linked Oligosaccharides 193 Fig Strategy of elucidation of oligosaccharide primary structure atoms; Gal H-3 and H-4 atoms; Fuc H-5 and H-6 ... presence of -2 , 3- or -2 ,6-linked sialic acid is clearly shown by the respective chemical shift of the H-3ax and H-3eq signals of the monosaccharides The H-3ax and H-3eq resonances of the -2 ,3-linked ... compared (compounds N-1 and A-1) 194 Table Chemical Shifts of GalNAc-ol Residues Characteristic of the Nature of Oligosaccharide-Alditol Cores Gal(β1–3)GalNAc-ol 194 H-2 H-3 H-4 H-5 H-6' 4.393 4.063...
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