Glycoprotein Methods and Protocols - P2

... 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-anti-gen ... mounting ,and its relative stability. There are, however, several other alternatives, such as 3-amino-9-ethylcarbazole (AEC), for use with horseradish peroxidase (HRP); 5-bromo-4-chloro-...

Ngày tải lên: 08/10/2012, 10:25

... al.for UDP-GlcNAc:GalNAc-Rβ1, 3-N-acetylglucosaminyltransferase (9). Actually, inin vitro experiments, these compounds inhibit the UDP-Gal:GalNAc-Rβ1,3-galac-tosyltransferase (8). GalNAcα-O-aryls ... with GalNAcα-O-aryl: The medium in ituma is added with aryl-N-acetyl-α-D-galactosaminide.c. Threonine-free medium (Life Technologies).d. Low-glucose Dulbecco’s minimal essential/H-16 medium (Gibc...

Ngày tải lên: 08/10/2012, 10:25

... H. (1986) Mucin synthesis: conversion of R 1- 1-3 Gal-R2to R 1- 1-3 (GlcNAcβ 1-6 )Gal-R 2and of R 1- 1-3 GalNAc-R2to R 1- 1-3 (GlcNAcβ 1-6 )GalNAc-R2by a β6-N-acetyl-glucosaminyltransferase in pig gastric mucosa. ... GlcNAcβ 1-6 (GlcNAcβ 1-3 ) GalNAcα-pnp, GlcNAcβ 1-3 Galβ 1-4 GlcNAc, GlcNAcβ 1-6 (GlcNAcβ 1-3 Galβ 1-3 ) GalNAcα-benzyl, GlcNAcβ 1-6 (GlcNAcβ...

Ngày tải lên: 08/10/2012, 10:25

... 100 mM IPTG; dissolve 23.8 mg isopropyl β-D-thiogalacto-pyranoside/mL in water. Filter sterilise. 2% X-gal: Dissolve 2% (w/v) 5-bromo-4-chloro-3-indoly-β-D-galactopyranoside in dimethyl formamide. ... β-galactosidase gene-coding region, allowing blue/white color screen-ing of clones with inserts in the presence of X-Gal and isopropyl-1-thiol-β-D-galactopyranoside (IPTG).Due to the large siz...

Ngày tải lên: 08/10/2012, 10:25

... expression bothin cell cultures (HT-29 MTX) and in tissues from various mucosae (trachea, bronchus,From :Methods in Molecular Biology, Vol. 125: Glycoprotein Methods and Protocols: The MucinsEdited by: ... Hadzopuolou-Cladaras, M., Nolte, R. T., Atkinson, D., and Zannis V.J.(1986) The complete sequence and structural analysis of human apolipoprotein B-100relationship between apo...

Ngày tải lên: 08/10/2012, 10:25

... integrate and orientate this mapin the general physical map of 11p15.5 including HRAS, D11S150, and IFG2 refer-ence markers.From :Methods in Molecular Biology, Vol. 125: Glycoprotein Methods and Protocols: ... regions of mammalian genomes, inPulsed-Field Gel Electrophoresis : Protocols, Methods and Theories, Methods in Molecu-lar Biology, vol. 12 (Burmeister, M., and Vlan...

Ngày tải lên: 08/10/2012, 10:25

... 0.1% Tween-20, pH 9.5.5. Nitroblue tetrazolium (NBT) salt.6. 5-bromo-4-chloro-3-indolyl phosphate toluidinium (BCIP) salt.2.9.2. Fluorescent ISH (FISH)1. Buffer 1: 0.15 M NaCl, 0.1 M Tris-HCl, pH ... useof tandem repeat (TR) probes to quantitate mRNA levels, especially with those mucingenes that exhibit polymorphisms.From :Methods in Molecular Biology, Vol. 125: Glycoprotein Methods and...

Ngày tải lên: 08/10/2012, 10:25

... sequenceaMUC1VNTRp 1-4 0 PDTRPAPGSTAPPAHGVTSAPDTRPAPGSTAPPAHGVTSAp 1-2 4 PDTRPAPGSTAPPAHGVTSAPDTRp 5-2 0 PAPGSTAPPAHGVTSAp1 3-3 2 PAHGVTSAPDTRPAPGSTAPC-p1 3-3 2 (C)PAHGVTSAPDTRPAPGSTAPp 1-1 5 PDTRPAPGSTAPPAHA-p 1-1 5 APDTRPAPGSTAPPAHN-terminal ... peptide: the peptide Cp1 3-3 2, derived from MUC1 VNTR region (contain-ing an N-terminal cysteine to form dimers); peptides from N-...

Ngày tải lên: 08/10/2012, 10:25

... glycosidases, e.g., α-sialidase, - and β-galactosidase (Oxford Glycosciences,Abingdon, UK), - and β-N-acetylhexosaminidase (Boehringer Mannehim), α-fucosidase(Sigma), O-glycanase (Calbiochem -Novabiochem).2. ... Protein biotinylation reagent: δ-biotinyl-ε-aminocaproic-N-hydroxysuccinimide ester(BNHS) (Sigma).4. Carbohydrate biotinylation reagent: δ-biotinyl-ε-aminocaproic-acid hydra...

Ngày tải lên: 08/10/2012, 10:25