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Laboratory TechniquesProtocolsScience
June 30, 2019

Ponceau S Stain Protocol

Ponceau S staining is a rapid and reversible staining method used for the detection of protein bands on Western blot membranes, Polyvinylidene fluoride (PVDF), nitrocellulose, and cellulose acetate membranes. Ponceau S is a negative stain that binds to the positively charged functional groups of the protein (amino group) and the…
Laboratory TechniquesProtocolsScience
June 30, 2019

Southern Blotting Protocol

Southern blotting is a hybridization technique used for deoxyribose nucleic acid (DNA). The method was named after Edward M. Southern, who developed the technique in the 1970s. The method involves the transfer of DNA fragments from an electrophoresis gel to a membrane causing immobilization of the DNA fragments and bands…
Laboratory TechniquesProtocols
June 26, 2019

Affinity Chromatography Protocol

Affinity chromatography is a versatile separation protocol that uses the biological interactions for characterization and detailed analysis of sample components. It is based on highly specific interactions between two molecules, such as the interactions between enzyme and its substrate, receptor, and ligand, or antibody and antigen. These reversible interactions are…
Laboratory TechniquesProtocols
June 26, 2019

Fast protein liquid chromatography (FPLC) Protocol

Fast protein liquid chromatography (FPLC) is a type of liquid chromatography that provides high resolution by small-diameter stationary phases for protein characterization and separation. The technique features high loading capacity, biocompatible buffer systems, fast flow rates, and stationary phases for common chromatography modes (e.g., gel filtration, ion exchange, reversed phase,…
Filtration Chromatography Protocol
Laboratory TechniquesProtocols
June 26, 2019

Gel Filtration Chromatography Protocol

Gel-filtration chromatography, also known as the size exclusion chromatography, is a versatile technique that permits the separation of proteins and other biological molecules. The gel filtration chromatography separates the proteins solely based on molecular size differences. For this, a porous matrix is used to which the molecules, for steric reasons,…
Laboratory TechniquesProtocols
June 26, 2019

Ion-exchange Chromatography Protocol

Ion Exchange Chromatography (IEC) is a powerful liquid chromatographic technique used for bioseparation. The separation is done by a reversible interaction between charged molecules of the sample with charged ligands attached to a column. The method offers a sizeable sample-handling capacity, powerful resolving ability, broad applicability, moderate cost, and the…
Laboratory TechniquesProtocols
June 26, 2019

KJELDAHL Method Protocol

The Kjeldahl method was developed by a brewer called Johann Kjeldahl in 1883. The protocol is built on the principle that strong acid helps in the digestion of food so that it releases nitrogen which can be determined by a suitable titration technique. By observing the nitrogen concentration of the…
Protein Quantification Protocols
Laboratory TechniquesProtocols
June 26, 2019

Protein Quantification Protocols

The estimation of proteins via the quantitative method is one of the basic requirements in biochemistry. Proteins, from various perspectives, are substantially more complex than nucleic acids. Thus, it has been hard to give laboratory protocols that can be applied to proteins. The precise quantitation of the amount of protein…