carbohydrate biotechnology protocols - christopher bucke
carbohydrate biotechnology protocols - christopher bucke
... Germany Document 页码， 1/1 http://emedia.netlibrary.com/nlreader/nlreader.dll?bookid=28014&filename=Page_1.html 200 3-5 -8 Page 1 1— Introduction to Carbohydrate Biotechnology Christopher Bucke The study of carbohydrates has been neglected in favor of consideration ... Production of 3-Keto-Derivatives of Disaccharides 277 Marie-Pierre Bousquet, René-Marc Willemot, Pierre F....
Ngày tải lên: 08/04/2014, 12:47
Glycoprotein methods protocols - biotechnology 048-9-001-013.pdf
... membrane-based meth- ods such as slot-blotting and staining with periodic acid-Schiff reagent; antibodies and lectins or coating methods such as the glycan detection method and enzyme-linked immunosorbent ... not seen with Sepharose gels. 2.4. Ion-Exchange High-Performance Liquid Chromatography Ion-exchange high performance liquid chromatography is carried out in our labora- tory using a Mono...
Ngày tải lên: 23/09/2012, 19:25
Glycoprotein methods protocols - biotechnology 048-9-027-044.pdf
... are, however, several other alternatives, such as 3-amino- 9-ethylcarbazole (AEC), for use with horseradish peroxidase (HRP); 5-bromo-4-chloro- 3-indolyl phosphate/nitro blue tetrazolium (BCIP/NBT); ... 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 eviden...
Ngày tải lên: 23/09/2012, 19:25
Glycoprotein methods protocols - biotechnology 048-9-045-055.pdf
... 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 chlo- ride mix for AP). Alternatively, ... nonionic detergents: 0.05% (v/v) Nonidet P-40 or Tween-20. 4. Enzyme substrates: 2,2'-azino-bis(3-ethylbenzathiazaline 6-sulfonic acid) (ABTS) (1 mg/mL, A 405nm ), O-phenyldiamine (OPD)...
Ngày tải lên: 23/09/2012, 19:25
Glycoprotein methods protocols - biotechnology 048-9-057-064.pdf
... Allen, A., Morris, E., and Ross-Murphy S. B. (1988) Mucus glycoprotein gels: role of glycoprotein polymeric structure and carbohydrate side-chains in gel-forma- tion. Carbohydrate Res. 178, 93–110. 7. ... water mixed with 250 mg of phenol in 70 mL of glycerol. 8. Poly- L -lysine-coated slides: slides are immersed in 0.01% (w/v) poly- L -lysine for 5 min and dried at room temperature. 3. M...
Ngày tải lên: 23/09/2012, 19:25
Glycoprotein methods protocols - biotechnology 048-9-065-073.pdf
... by Western-type dot-blot procedure. Spot aliquots of the homogenates or media on nitrocel- lulose paper, using the dot-blot apparatus and air-dry for 5 min. Perform all ensuing pro- cedures in ... equiv- alent apparatus) to quantify the amount of radioactivity in an identified band on SDS-PAGE. 6. Poly- or monoclonal antibodies (IgG-type) against nonglycosylated regions of the mucin- polypept...
Ngày tải lên: 23/09/2012, 19:25
Glycoprotein methods protocols - biotechnology 048-9-075-085.pdf
... assessment of pep- tide homogeneity is advisable. We analyze aliquots of the peaks by using matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS), and purify ... with “naked” cysteine-rich domains (4–7). Histological and biochemical investigations suggested that mucous-producing tis- sues and their secretions contained a complex mixture of mucin-type glyc...
Ngày tải lên: 23/09/2012, 19:26
Glycoprotein methods protocols - biotechnology 048-9-087-096.pdf
... molecules not on hydrodynamic volume alone but on their mass-to-volume ratio as well. The basis of rate-zonal centrifugation for this pur- pose is long established and, in its simplest form, is ... Distribution of Gel-Forming Mucins John K. Sheehan and David J. Thornton 1. Introduction The rheological properties of human mucus are dominated by the physical proper- ties of large secreted O-lin...
Ngày tải lên: 23/09/2012, 19:26
Glycoprotein methods protocols - biotechnology 048-9-111-119.pdf
... 9-fluorenylmethyl oxycarbonyl chloride (FMOC )- based precolumn derivatization amino acid analysis that has been extensively vali- dated (1,2). Although the detailed protocols on the use of the automated ... mucin gly- coproteins. 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 reve...
Ngày tải lên: 23/09/2012, 19:26
Glycoprotein methods protocols - biotechnology 048-9-121-128.pdf
... window with elution profiles of PTH-threonine-GalNAc-Gal (black arrow- heads) and PTH-threonine-GalNAc (gray arrowheads) from the Applied Biosystems Procise. Stan- dard amino acids are shown for ... from Edman degrada- tion: the identification of Xaa-Pro-Xaa-Xaa as a motif for Thr-O-glycosylation. Biochem. Biophys. Res. Commun. 178, 1194–1201. 6. Laursen, R. A. (1971) Solid-phase Edman degrad...
Ngày tải lên: 23/09/2012, 19:26