A Method for Micro-Scale Isolation and Purification of Gangliosides Solvent partition protocol allows the isolation of gangliosides from small samples and from samples where ganglioside concentrations are low, especially relative to the concentration of potentially contaminating proteins and other large molecular weight species. Stephan Ladisch Director, Center for Cancer and Transplant Biology, Children's National Medical Center, Washington, D.C.
A Sensitive ELISA-Based Assay for Glycosyltransferases The method allows the detection and quantification of glycosyltransferase activity using an ELISA-based procedure and carbohydrate-specific monoclonal antibodies. Avoids the use of radiolabeled substrates. Bruce A. Macher~Professor of Chemistry and Biochemistry, San Francisco State University, San Francisco, CA
ABH (p-Azidobenzoyl hydrazide) PDF The ABH crosslinker binds to the Fc portion of an antibody molecule, away from the antigen binding site, resulting in a
divalent immunologically active immunoglobulin derivative. Pierce
Analysis of Oligosaccharide Ligands by High Performance Liquid Affinity Chromatography High performance liquid affinity chromatography (HPLAC) is a useful procedure to investigate he interactions between carbohydrate binding protein and their ligands. Technical requirements are similar to conventional HPLC. HPLAC can screen and separate natural ligands from complex biological mixtures. WeiTong Wang~GlycoTech Corporation, Rockville, Maryland
Carbohydrate Assays o determine the relative amounts of
LPS carbohydrates present in a given strain. The assay can be done on one set of samples and then scanned at the various wavelengths for reasonable data on the 3 types of sugars.
HEXOSE ASSAY, 6-DEOXYHEXOSE ASSAY, HEPTOSE ASSAY. Hancock Laboratory.
Deglycosylation of Glycoproteins Using Endoglycosidases Deglycosylation steps are different with every glycoprotein and must be determined empirically (Methods in Enzymology, 1994, 230: 44-57). A typical test reaction of human transferrin is described.
T.H. Plummer, Jr. and A.L. Tarentino, Department of Biochemistry, New York State Department of Health, Wadsworth Center for Laboratories and Research, Albany, New York
Immunohistochemistry using Anti-Ganglioside Antibodies Indirect method measuring immunofluorescence coupled to second antibody. Best for membrane antigens in addition to intra- and extracellular antigens, may be applied to frozen tissue sections, to cells in suspension, and to cells attached to glass slides or coverslips. Tadashi Tai~Head, Department of Tumor Immunology, The Tokyo Metropolitan Institute of Medical Science, Tokyo, Japan
Immunostaining Thin Layer Chromatograms Of Glycolipids Immunostaining thin layer chromatograms TLC is a very sensitive detection technique of functionally active carbohydrate ligands of protein receptors. Carbohydrate structures are detected in glycolipids from complex mixtures of molecules extracted from the relevant target tissue. Proteins analyzed can be antibodies, chimeric Ig proteins, selectins, lectins, toxins, and other carbohydrate binding proteins. John L. Magnani~GlycoTech Corporation, Rockville, Maryland
PDP-Hydrazide PDF PDP-Hydrazide 3-(2-pyridyldithio)propionyl hydrazide is heterobifunctional cleavable crosslinker containing a hydrazide group and pyridyl disulfide residue that react with carbohydrates and sulfhydryls, respectively.