title: Solubilization of proteins: the importance of lysis buffer choice.
abstract: The efficient extraction of proteins of interest from cells and tissues is not always straightforward. Here we demonstrate the differences in extraction of the focal adhesion protein Kindlin-2 from choriocarcinoma cells using NP-40 and RIPA lysis buffer. Furthermore, we demonstrate the use of a more denaturing urea/thiourea lysis buffer for solubilization, by comparing its effectiveness for solubilization of small heat-shock proteins from smooth muscle with the often utilized RIPA lysis buffer. Overall, the results demonstrate the importance of establishing the optimal lysis buffer for specific protein solubilization within the experimental workflow.
authors.affiliation: Division of BioMedical Sciences, Health Sciences Centre, Rm 5335, 300 Prince Philip Drive, St. John's, NL, Canada, A1B 3V6.
authors.forename: Mandy
authors.initials: M
authors.lastname: Peach
authors.forename: Noelle
authors.initials: N
authors.lastname: Marsh
authors.forename: Ewa I
authors.initials: EI
authors.lastname: Miskiewicz
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: MacPhee
mesh_terms: D000818:Animals; D002021:Buffers; D045744:Cell Line, Tumor; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D015151:Immunoblotting; D047468:Immunoprecipitation; D008565:Membrane Proteins / Q000032:analysis / Q000737:chemistry* / Q000302:isolation & purification; D008567:Membranes, Artificial; D017830:Octoxynol; D011092:Polyethylene Glycols / Q000737:chemistry; D012995:Solubility
chemical_list: D002021:Buffers; D008565:Membrane Proteins; D008567:Membranes, Artificial; D011092:Polyethylene Glycols; D017830:Octoxynol; C010615:Nonidet P-40