Polyvinylidene difluoride membranes

The separated proteins were transferred to a polyvinylidene difluoride membrane, and nonspecific IgC binding sites were blocked by incubation with 5% nonfat dry milk for 1 h at room temperature. The membranes were then incubated overnight at 4°C with primary antibodies. Immune complexes were detected by enhanced chemiluminescence (Amersham Biosciences). 

Use of Proteins Blotted to Polyvinylidene Difluoride Membranes as Immunogens... 

Matsudaira, P. (1987) Sequence from picomole quantities of proteins electroblotted onto polyvinylidene difluoride membranes J. Biol. Chem. 262, 10,035-10,038. 

Wise, G. E. and Lin, F. (1991) Transfer of silver-stained proteins from polyacrylamide gels to polyvinylidene difluoride membranes. J. Biochem. Biophys. Methods 22, 223-231. 

Masuoka, J., Glee, P.M., and Hazen, K.C. (1998). Preparative isoelectric focusing and preparative electrophoresis of hydrohobic Candida albicans cell wall proteins with in-line transfer to polyvinylidene difluoride membranes for sequencing. Electrophoresis 19, 675-678. 

The sample was separated by SDS-PAGE (Laemmli, 1970) (Multigel 4/20, Daiichi Pure Chemicals Co., Ltd., Tokyo, Japan) and the separated proteins were subsequently electroblotted onto a polyvinylidene difluoride membrane (Immobilon, Millipore Corporation, Bedford, MA, USA). Antigens were probed with HIV-1-positive plasma and 0.5beta (a kind gift from Dr. Shuzo Matsushita of Kumamoto University, AIDS Research Institute, Kumamoto, Japan). The bands were visualized by means of chemiluminescence detection (NEN Life Science Products, Inc., Boston, MA, USA). 

Kennedy, T., et al. (1988). Sequencing of Proteins from Two-Dimensional Gels by Using In Situ Digestion and Transfer of Peptides to Polyvinylidene Difluoride Membranes Application to Proteins Associated with Sensitization iriAplysia Proc. Natl. Acad. Sci. USA 85 7008-7012. 

Ogawa, H., et al. (1990). Direct Carbohydrate Analysis of Glycoproteins Electroblotted onto Polyvinylidene Difluoride Membrane from Sodium Dodecyl Sulfate Polyacrylamide Gel, Anal. Biochem. 190 165-169. 

I.F. Wang, J.F. Ditter, R. Zepf (USF Filtration and Separations) Highly porous polyvinylidene difluoride membranes. US Patent 5834107, November 1998. 

Isobe, T., Naiki, M., Handa, S. and Taki, T. (1996) A simple assay method for bacterial binding to glycosphingolipids on a polyvinylidene difluoride membrane after thin-layer chromatography blotting and in situ mass spectrometric analysis of the ligands. Anal Biochem., 236, 35-40. 

TLC/immunostaining has been used to detect solaso-dine glycosides by separation of the compounds on a silica gel layer, transfer to a polyvinylidene difluoride membrane, and treatment of the membrane with sodium periodate solution followed by bovine serum albumin (BSA), resulting in a solasodine-BSA conjugate. Individual zones were stained by monoclonal antibody against solamargine. 

Bottino A., Roda G.C., CapanneUi G., Munari S.M. (1991),The formation of micro-porous polyvinylidene difluoride membrane by phase separation, / Membrane Sci., 57,1-20. 

Taki, T., Ishikawa, D., Handa, S., and Kasama, T. (1995). Direct mass spectrometric analysis of glycosphingolipid transferred to a polyvinylidene difluoride membrane by thin layer chromatography blotting. Anal. Biochem. 225 24-27. 

Nitrocellulose or polyvinylidene difluoride membranes are often utilized for western blotting. Because of their moderate levels of protein retention and comparably low price, nitrocellulose membranes have been well accepted for basic detection. However, polyvinylidene difluoride membranes possess better protein retention, physical strength, and chemical resistance. Thus they are often used when reprobing or protein sequencing will be performed because they will not be affected by repeated washing with organic solvents. 

Ishikawa, D. and Taki, T. 2000. Thin-layer chromatography blotting using polyvinylidene difluoride membrane (Far-Eastern blotting) and its apphcations. Sphingolipid Metabolism and Cell Signaling Pt B, 312 145-57. 

Bottino, A., G. Camera-Roda, G. Capannelli, and S. Munari. 1991. The Formation of Microporous Polyvinylidene Difluoride Membranes by Phase Separation. Journal of Membrane Science 57(1) 1-20. doi 10.1016/S0376-7388(00)81159-X. 

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