"Discover the ins and outs of polyacrylamide gel electrophoresis (PAGE) with detailed insights on applications, procedures, and requirements. Learn about sample preparation, gel preparation, electrophoresis steps, and various applications such as molecular weight measurement, protein size estimation, and more."
Please find below an Image/Link to download the presentation.
The content on the website is provided AS IS for your information and personal use only. It may not be sold, licensed, or shared on other websites without obtaining consent from the author. If you encounter any issues during the download, it is possible that the publisher has removed the file from their server.
You are allowed to download the files provided on this website for personal or commercial use, subject to the condition that they are used lawfully. All files are the property of their respective owners.
The content on the website is provided AS IS for your information and personal use only. It may not be sold, licensed, or shared on other websites without obtaining consent from the author.
E N D
Presentation Transcript
Electrophoresis Part II Dr. Shalini Verma BBT-201 Department of Biotechnology IBSBT, CSJM, University, Kanpur
Polyacrylamide Gel Electrophoresis cross-linked gels formed by the polymerization of acrylamide with a cross-linking agent, usually N,N -methylenebisacrylamide. free radical polymerization ammonium persulfate as the initiator and N,N,N ,N -tetramethylethylendiamine (TEMED) as the catalyst. widely used in biochemistry, forensic chemistry, genetics, molecular biology and biotechnology to separate biological macromolecules, usually proteins or nucleic acids, according to their electrophoretic mobility. commonly used form of polyacrylamide gel electrophoresis is the Sodium dodecyl suplhate Polyacrylamide gel electrophoresis (SDS- PAGE)
Requirements for Polyacrylamide Gel Electrophoresis (PAGE) Acrylamide solutions (for resolving & stacking gels). Isopropanol / distilled water. Gel loading buffer. Running buffer. Staining, destaining solutions. Protein samples Molecular weight markers. The equipment and supplies necessary for conducting SDS-PAGE includes: An electrophoresis chamber and power supply. Glass plates (a short and a top plate). Casting frame Casting stand comb
Applications of Polyacrylamide Gel Electrophoresis (PAGE) Measuring molecular weight. Peptide mapping. Estimation of protein size. Determination of protein subunits or aggregation structures. Estimation of protein purity. Protein quantitation. Monitoring protein integrity. Comparison of the polypeptide composition of different samples. Analysis of the number and size of polypeptide subunits. Post-electrophoresis applications, such as Western blotting. Staining of Proteins in Gels with Coomassie G-250 without Organic Solvent and Acetic Acid. Pouring and Running a Protein Gel by reusing Commercial Cassettes. Selective Labelling of Cell-surface Proteins using CyDye DIGE Fluor Minimal Dyes. Detection of Protein Ubiquitination.
