title: Effective solubilization procedure for analysis of silkworm hemolymph proteins by two-dimensional gel electrophoresis.
abstract: The outstanding capability of two-dimensional gel electrophoresis in separating all types of proteins basically depends on the efficiency of sample preparation. Sample preparation is one of the most critical steps in two-dimensional gel electrophoresis. Unfortunately, due to severe solubility, resolution of protein on gel is usually hampered, and thus, analysis remains a difficult task. However, technically several problems are generally encountered during protein extraction and isoelectric focusing. In the present investigation, we emphasized on evaluation and comparison of six different protein solubilization methods intended for resolving and analyzing silkworm hemolymph proteins by two-dimensional gel electrophoresis. Our findings revealed that the buffer composition of 8 M urea, 4 % 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate, 40 mM Tris base, 65 mM dithiothreitol, and 0.2 % ampholyte can effectively solubilize and yields maximum protein spots.
authors: Department of Sericulture, Karnatak University, Dharwad, 580 003, Karnataka, India. gspmalli@gmail.com
authors: Mallikarjun
authors: M
authors: Gadwala
authors: Neetha
authors: N
authors: Kari
authors: Narayan
authors: N
authors: Moger
authors: Shyam Kumar
authors: SK
authors: Vootla
mesh_terms: D000818:Animals; D012831:Bombyx / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D006458:Hemolymph / Q000378:metabolism*; D019476:Insect Proteins / Q000378:metabolism*; D055550:Protein Stability; D012995:Solubility
chemical_list: D019476:Insect Proteins