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]
Solutions for Agarose gel prepartion, Selection of Appropriate Gel Size and Composition. Mycology Lab
University of Tennessee - [Read Agarose Gel Preparation Protocol]
An introduction to AFLP and fAFLP. Mark E. Berres, University of Wisconsin. Amplified fragment-length polymorphism (AFLP) or its fluorescent version (fAFLP) is a PCR-based fingerprinting technology. AFLP basically involves the restriction of genomic DNA - [Read An introduction to AFLP and fAFLP]
An excellent guide on the analysis of proteins on SDS-PAGE gels, through staining with coomassie blue dye and western blot analysis. Analysis of Protein Gels (SDS-PAGE). David R. Caprette, Rice University. - [Read Analysis of Protein Gels (SDS-PAGE)]
Arabidopsis transformation and selection on kanamycin agar, Preparation of plants for transformation, Agrobacterium, Dipping, Post-dip care, Agar plates, Tips. Adaptation of the short protocol described bySteve Clough and Andrew Bent, University of Illinois at Urbana-Champaign. - [Read Arabidopsis transformation with pictures]
Adapted from Frommer et.al. Good protocol for bisulfite treatment of DNA. Includes tips on Methylation PCR for CpG methylation analysis. University of Texas M. D. Anderson Cancer Center - [Read Bisulfite Treatment of DNA for Methylation Analysis]
Blocking of unwanted non-specific staining in Immunofluorescence. Blocking of endogenous enzymes, Blocking of endogenous fluorochromes, Blocking endogenous biotin , Blocking of endogenous Fc blocking, Blocking of crossreactive antigens in the tissue. Cattoretti. Columbia University - [Read Blocking of unwanted non-specific staining in Immunofluorescence.]
Protocol detects specific cell adhesion to glycolipids resolved on TLC plates. Carbohydrate-Specific Adhesion of Intact Cells to Resolved Glycolipids on TLC Plates. Ronald L. Schnaar~Professor, Johns Hopkins University Medical School, Baltimore, Maryland. Glycotech. - [Read Carbohydrate-Specific Adhesion of Intact Cells to Resolved Glycolipids on TLC Plates]
cDNA Library Screening Protocol - ZAPII Detailed Protocol. XLI-Blue MRF' and cDNA phage library screening. Dr. Laurie. University of Virginia Health Science Center - [Read cDNA Library Screening Protocol - ZAPII]
PCR Reaction Conditions for Colony PCR. Contributed by Lynn Hancock. Enterococcus Research Site, The Board of Regents of the University of Oklahoma. - [Read Colony PCR Protocol]
SAVING OF DAPI-IMAGES, SLIDE PRETREATMENT, PROBE PREPARATION, PROBE DETECTION, washing blocking detection, Counterstaining with DAPI. Institute of Pathology,Humboldt-University of Berlin - [Read COMPARATIVE GENOMIC HYBRIDIZATION (CGH) PROTOCOL]
Concentration of DNA by Ethanol Precipitation Protocol. Adapted fromBruce A. Roe, Department of Chemistry and Biochemistry, The University of Oklahoma, Norman, Oklahoma. Usually 2.5 - 3 volumes of ethanol and/or acetate solution is added to the DNA in a microcentrifuge tube. This is then put into an ice-water bath for at least 10 minutes. The precipitation is performed by incubation at -20C overnight. - [Read Concentration of Oligo DNA by Ethanol Precipitation Protocol]
DNA digestion protocol & hints. Procedure and tips for restriction digestion. Doering laboratory, Department Molecular Microbiology, Washington University Medical School. - [Read DNA digestion protocol & hints PDF]
DNA Extraction from Agarose Gels Protocol. The page includes cutting out the DNA band from the gel, and describes three methods including 1) Spin-columns (Nucleic acid purification columns), 2) using Dialysis tubing (semi-permeable membrane, Visking tubing), and the 3) Paper strip method.Matt Lewis, Department of Pathology, University of Liverpool. - [Read DNA Extraction from Agarose Gels 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.
In Vitro Translated Xenopus Mos Kinase Assay Protocol. In response to progesterone, immature Xenopus oocytes mature to eggs that can be fertilized. The Mos protein kinase is essential for oocyte maturation, most likely due to its ability to activate the MAP kinase cascade. This MAP kinase cascade eventually leads to the activation of Cdc2/cyclin B and entry into M phase. In this protocol, tagged Mos kinase is translated in vitro, immunopurified, and used in a kinase assay.
A protocol for the selection of Phage Antibodies using Immobilized Antigen. This method describes the selection of antibodies from bacteriophage antibody libraries that recognize a specific antigen. The phage display library of antibody-displaying phage particles is exposed to antigen attached to a solid substrate (Nunc Immuno™ tubes). The phage particles with affinity for antigen bind to the immobilized antigen and are selected from the library of phage expressing antibodies.