... 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, including acetylcholinesterase, 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,...
... such as Culling’s Handbook ofHistopathological and Histochemical Techniques (including museum techniques) (36) orKiernan’s Histological and Histochemical Methods: Theory and Practice (45) providecomprehensive ... 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...
... Chemical, Lectin, and Antibody Methods Michael A. McGuckin and David J. Thornton1. IntroductionDetection and quantitation of mucins can be important in both the research and clinical settings. ... 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 ... sample volume, and superior wash-ing). For quantitation and comparison across blots, a standard in the same buffer assamples should be titrated for use as a standard curve, and samples should...
... poly-L-lysine for 5 min and dried at room temperature.3. Methods 3.1. Observation and Measurementof Mucous Thickness on Unfixed Mucosal Sections (see Notes 1–4)This is a rapid and simple method ... will be greatlyreduced and discontinuous.7. Careful handling of the cryostat sections on the slide during fixing and staining is impor-tant because excessive washing and so on can remove mucus ... to rat, frog and human gastric mucosa: influence of feedingprostaglandin, N-acetyl-cysteine and other agents. Clin. Sci. 63, 187–195.4. Bell, A. E., Allen, A., Morris, E. R., and Ross-Murphy,...
... samples no. 1 and 2 (values a and e):x = g/c ì a/e (no dimension) Biosynthesis and Secretion of Mucin 6565From: Methods in Molecular Biology, Vol. 125: Glycoprotein Methodsand Protocols: ... 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...
... small gel, use 1.6 g ofagarose and 160 mL of buffer, and for a large gel, use 2.8 g of agarose and 280 mL ofbuffer.2. Leave to cool before pouring (hand hot) and insert well-forming comb (see ... 1.5 mm thick and 1 cm wide. There is a compromise betweenband broadness and amount of sample to be loaded. Larger amounts of sample tend toyield poorer quality data; i.e., the bands become more ... other identified mucins. Separation and Identification of Mucin 777Separation and Identificationof Mucins and Their GlycoformsDavid J. Thornton, Nagma Khan, and John K. Sheehan1. IntroductionThis...
... 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 contrast, including ... 2.400aThe 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: ... 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...
... 113113From: Methods in Molecular Biology, Vol. 125: Glycoprotein Methodsand Protocols: The MucinsEdited by: A. Corfield â Humana Press Inc., Totowa, NJ10Amino Acid Analysis of MucinsJun X. Yan and ... 2 min for pump and column equilibration with the initial composition ofthe mobile phase.5. Figures 1 and 2 show typical chromatograms of the separation of amino acid standards and hydrolyzed ... samples and Dr. Andrew Gooley and Prof. Keith Williams for their support.References1. Yan, J. X., Wilkins, M .R., Ou, K., Gooley, A. A., Williams, K. L., Sanchez, J C.,Golaz, O., Pasquali, C., and...
... 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. ... 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 ... 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 àg 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....