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 - [Read A Sensitive ELISA-Based Assay for Glycosyltransferases]
Restriction and Ligation reactions, Setting up the reactions, AFLP PCR reactions protocols. Analyzing the data using Genescan. Paul G. Wolf, Utah State Univ. - [Read AFLP protocol Wolf Lab]
Electrotransformation of Agrobacterium with a plasmid that has been replicating in E. coli. Growth of Arabidopsis thaliana. Arabidopsis dunking. Seed Harvesting. Plant tissue culture. Very detailed protocol. Stockinger lab. PDF - [Read Arabidopsis transformation with Agrobacterium PDF]
There are two basic methods for the in vitro assay of B-galactosidase activity from yeast. They
differ mainly in the method of preparing the material for assay. Both methods are described with accompanying protocols. Method I: Assay of Crude Extracts includes: Yeast Cell Growth; Yeast Cell Harvest; B-gal assays; Bradford Assays. Method II: Permeabilized cell assay. - [Read Assay of β-Galactosidase in Yeast Protocol]
Centrifuge Safety - High Speed Hazards. Safe Centrifuge Use , Care and Maintenance, and maintaining a log book for centrifuges. EHS Washington state Univ. - [Read Centrifuge Safety - High Speed Hazards]
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 - [Read Deglycosylation of Glycoproteins Using Endoglycosidases]
Fixation is to preserve cells and tissue constituents in as close a life-like state as possible and to allow them to undergo further preparative procedures without change. Fixation arrests autolysis and bacterial decomposition and stabilises the cellular and tissue constituents so that they withstand the subsequent stages of tissue processing. Great detailed guide with protocols. Anthony S-Y Leong. - [Read Fixation and Fixatives - A great guide]
There are many variations based on the type of tissue to be examined, whether one is dealing with cell suspensions, biopsies, perfused tissues, or monolayer . Fixation Protocol. OSU Campus Microscopy & Imaging Facility :: The Ohio State University College of Med - [Read Fixation Protocol]
Isolation of DNA fragments using glass milk (GENE-CLEAN). Glass Milk Agarose Gel DNA Extracton Protocol. Minion Lab, College of Veterinary Medicine at Iowa State University. - [Read Glass Milk Agarose Gel DNA Extracton Protocol]
Protocol for GUS reporter gene assay. Includes: Protein isolation; Alternative method for small (<1g) quantities of tissue; GUS assays; Bradford protein concentration determination assays - [Read GUS Reporter Gene Assay Protocol]
Tubulin is polymerized into microtubules by incubating tubulin at 37°C with GTP. A nucleation seed is added when the purpose is to assay microtubule elongation. Tubulin can also be polymerized for the purposes of recycling the tubulin or labeling the microtubules with fluorescently labeled tubulin. Based on the protocol by Timothy Mitchison of Harvard University.