... UV monitor and collect fractions using a fraction col-lector and subject to the appropriate carbohydrate and antibody analyses.3.6. Analysis of Mucins Methods for the detection and analysis ... antibodies and lectins or coating methods such as the glycan detection method and enzyme-linkedimmunosorbent assays (ELISA). All these techniques have advantages and disadvan-tages. Solution methods ... serine proteases and esterases, includingacetylcholinesterase, and should therefore be handled in a fume cupboard with extremecare! DFP is supplied in 1-g vials with a septum, and prior to dilution,...
... Rhodes, J. M. and Milton, J. D. (1986) Lectin Methodsand Protocols, Humana, Totowa,NJ.50. Liener, I. E., Sharon, N., and Goldstein, I. J. (1998) The Lectins: Properties, Functions, and Applications ... nature and limitations of the individual techniques, and (2) interpretation and reporting of mucinstaining.1.1. Nature and Limitations of Mucin-Staining Methods Mucin histochemistry, lectin, and ... such as Culling’s Handbook ofHistopathological and Histochemical Techniques (including museum techniques) (36) orKiernan’s Histological and Histochemical Methods: Theory and Practice (45) providecomprehensive...
... in Glycoprotein Methods and Protocols: The Mucins (Corfield, T., ed.), Humana, Totowa, NJ.3. Carraway, K. L. (2000) Preparation of membrane mucin, in Glycoprotein Methods and Protocols: The Mucins ... Nagma, K., Thornton, D. J., Khan, N., and Sheehan, J. K. (2000) Separation and identifi-cation of mucins and their glycoforms, in Glycoprotein Methodsand Protocols: TheMucins (Corfield, T., ... 1 and 2 (2,3) of this volume for the preparation ofsecreted and membrane-associated mucins, respectively, and to Chapter 7 (4) for adiscussion of methods for mucin separation.50 McGuckin and...
... samples no. 1 and 2 (values a and e):x = g/c × a/e (no dimension)Biosynthesis and Secretion of Mucin 6565From: Methods in Molecular Biology, Vol. 125: Glycoprotein Methodsand Protocols: The ... PhosphorImager.4. The identification of mucin precursors and mature mucins using polypeptide-specificantisera and SDS-PAGE is elaborated in Chapters 20 and 21, and in several references(1,4–11). The precursor ... NJ6Quantitation of Biosynthesis and Secretion of Mucin Using Metabolic LabelingJan Dekker, B. Jan-Willem Van Klinken,Hans A. Büller, and Alexandra W. C. Einerhand1. IntroductionMost epithelial...
... values for ρ and ηrel. are from ref. 14.Heterogeneity and Size Distribution of Gel-Forming Mucins 8787From: Methods in Molecular Biology, Vol. 125: Glycoprotein Methodsand Protocols: The ... variety of different sizes and should be chosen according tosample volume and concentration.Heterogeneity and Size Distribution of Gel-Forming Mucins 91mucins on these grids; and the addition of ... K. E., and Chapman J. A. (1985) Mica sandwhichtechnique for preparing macromolecules for rotary shadowing. J. Ultrastruct. Res. 91,66–76.14. Kawahara K and Tanford C. (1966) Viscosity and density...
... Curmi, P. M. G., Williams, K. L., and Gooley, A. A. (1996)Sequencing Glycopeptides 121121From: Methods in Molecular Biology, Vol. 125: Glycoprotein Methodsand Protocols: The MucinsEdited by: ... Kirk, M., Barnes, S., Lowary, T. L., and Hart, G.W. (1996) Selective detection and site-analysis of O-GlcNAc-modified glycopeptides byβ-elimination and tandem electrospray mass spectrometry. ... peptides.Acta. Chem. Scand. 4, 283–293.4. Edman, P and Begg, G., (1967) A protein sequencer. Eur. J. Biochem. 1, 80–91.5. Gooley, A. A., Classon, B. J., Marschalek, R., and Williams, K. L. (1991)...
... lysates and media. Nonspecificbands may be present and can be eliminated from consideration using the pSVL vectorcontrol lanes for comparison (Fig. 2A,B, lanes 1 and 3). The sizes of specific bandsreported ... polymerase, and incubate the reaction at 72˚C to allow extension of the linkedtemplates. Add 2 µg each of primers 5 and 6, and continue the PCR with 30 cycles at 94˚Cfor 30 s, 50˚C for 1 min and 72˚C ... 195–205.12. Bhargava, A. K., Woitach, J. T., Davidson, E. A., and Bhavanandan, V. P. (1990) Cloning and cDNA sequence of a bovine submaxillary gland mucin-like protein containing twodistinct domains....
... (Neu5Gc), and keto-deoxy-nonulosonic acid (Kdn), respectively. Substituents of the hydroxyl groups present onFrom: Methods in Molecular Biology, Vol. 125: Glycoprotein Methodsand Protocols: ... Nakamura, M., and Ohkura, Y. (1987) Fluoro-metric high performance liquid chromatography of N-acetyl- and N-glycolyl neuraminicacids and its application to their microdetermination in human and animal ... at 2°C/min.5. Use injector and detector temperatures of 240 and 250°C, respectively.174 Michalski and CaponTable 4Elution Gradient for the Separationof Unreduced and Reduced MonosaccharidesTime...
... M. J. and Hounsell, E. F. (1998) HPLC and HPAEC of oligosaccharides and glyco-peptides, in Glycoscience Protocols (Hounsell, E. F., ed.), Humana, Totowa, NJ, pp. 79–100.14. Kakehi, K., and Honda, ... 0.5 cm column) and elute with threecolumn vol of HPLC-grade water.5. Collect the effluent and eluent and then pool (2.5-mL volume).6. Load onto a PD10-Sephadex GM25 column and elute with HPLC-grade ... in Glycoanalysis Protocols, vol. 76 (Hounsell, E.F., ed.), Humana Press, Totowa, NJ.21. Greis, K. and Hart G. (1998) Analytical methods for the study of O-GlcNAc glycoproteins and glycopeptides,...
... or three atthe most) and the facilities that are available in the laboratory environment will defineFrom: Methods in Molecular Biology, Vol. 125: Glycoprotein Methodsand Protocols: The MucinsEdited ... carbons 4 and 5 and also the exocyclic chain ofsialic acid when it is not 8 or 9 substituted. The oligosaccharide-alditol is cleaved intwo fragments, one corresponding to the 3-branch and the ... was introduced in the 1970s and rapidlyreceived a large application for analyzing the sequence of N-acetyllactosamine- and oligomannosidic-type glycans.Originally, and before the development...