Polyacrylamide Gel Electrophoresis: An Analytical Technique to Detect Heparan Sulfates of Varying Chain Lengths Isolated from Mouse Lung Tissue

Sulfated glycosaminoglycans like heparan sulfates, HSs, are negatively charged, linear, sulfated polysaccharides of variable chain lengths, attached to cell surface proteins.

To estimate different-length HSs isolated from mouse lung tissue, begin with gel cassette assembly. Partially fill it with highly concentrated acrylamide-bisacrylamide solution, including catalyzing agents - ammonium persulfate and tetra-methyl-ethylene-diamine. Overlay with water to prevent air contact, which may inhibit polymerization. Incubate.

Catalyzing agents induce acrylamide-bisacrylamide polymerization, forming small-pore resolving gel. Discard water. Add low concentration of acrylamide-bisacrylamide solution with catalyzing agents. Insert a gel comb to create wells. The solution polymerizes, forming large-pore stacking gel.

Load the electrophoresis tank's chambers with running buffer for optimum electrical conductivity during electrophoresis.

Dissolve the HS mixture in loading buffer containing tracking dye and sucrose. Next, transfer this mixture and standard HS ladders into cassette wells. Sucrose confines the mixture to the wells. Dye helps monitor electrophoresis progression.

Initiate electrophor