biochemistry Protocols

Category: Molecular Biology Protocols: Carbohydrate Protocols

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]

Category: Molecular Biology Protocols: Agarose Gel Preparation

AGAROSE GEL PROCEDURE in pictures. Biochemistry at St. Olaf College Muth Lab - [Read AGAROSE GEL PROCEDURE]

Category: Protein Protocols: Protein Quantitation Concentration Protocols: Bradford Protein Assay Protocols

The bradford dye-binding assay is a colorimetric assay for measuring total protein concentration.  It involves the binding of Coomassie Brilliant blue to protein. There is no interference from cations nor from carbohydrates such as sucrose. 
Detergents such as sodium dodecyl sulfate and triton x-100 can interfere with the assay, as well as strongly alkaline solutions.

Includes a general overview of the procedure and preparation of the standards in the protocol. - [Read Bradford Assay Method]

Category: Chromatography Protocols: Column Chromatography Protocols

Size Exclusion Column Chromatography Protocol. PDF. In this protocol you will learn how to use three types of column chromatography: Gel Filtration or Size Exclusion (SEC), Ion Exchange (IEC), and affinity (AC) in order to purify proteins and enzymes based on the physical properties of these biomolecules. Univ. Arizona, Biochemistry. - [Read Column Chromatography Protocol]

Category: Oligonucleotide Protocols: Oligonucleotide Purification Protocols

Concentration of DNA by Ethanol Precipitation Protocol. Adapted from Bruce 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]

Category: Molecular Biology Protocols: Carbohydrate Protocols

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]

Category: ELISA Protocols

ELISA (Enzyme-Linked ImmunoadSorbent Assay) Method and technique. Jill Fergusson, Department of Biochemistry, University of Nottingham, - [Read ELISA (Enzyme-Linked ImmunoadSorbent Assay)]

Category: General Research Protocols and Methods: Sterile Technique

Formaldehyde sterilization of tissue culture hoods. A bit toxic however effective. Dr.Dawson, Department Biochemistry, Univ. Nottingham Medical School. - [Read Formaldehyde Treatment of Tissue Culture Hoods]

Category: DNA Protocols: DNA Extraction Protocols

Large scale double-stranded DNA isolation, Midiprep double-stranded DNA isolation. Bruce A. Roe, Department of Chemistry and Biochemistry, The University of Oklahoma, Norman. - [Read Methods for DNA isolation - Protocol Book]

Category: ELISA Protocols: Sandwich ELISA Assay Protocols

Sandwich ELISA Procedure. Kitto Lab, Univ. Texas at Austin - [Read Sandwich ELISA Procedure]