Detection of heat shock protein-70 from Trichinella spiralis larvae using a modification of the routine western blotting procedure.
This is the first study that establishes a standardized western blotting method for the detection of heat shock protein (HSP) 70 from Trichinella spiralis using (selected) monoclonal antibodies (mAbs). Enhancement of HSP transfer onto the supportive membrane and increased retention of protein by the membrane are prominent features of the procedure. The reactivity of T. spiralis HSP70 on western blots was substantially increased by the use of a 10% acrylamide gel, the optimization of conditions during electrotransfer, and transfer onto Immobilon membranes. These data indicate that mAbs actually capable of detecting the agent of interest may be discarded because of nonoptimal testing conditions. We suggest that this method will aid in understanding the role and function of T. spiralis HSP70 in host-parasite (inter)relationship(s).
Martinez
J
J
Department of Microbiology and Parasitology, Faculty of Pharmacy, University of Alcala, Madrid, Spain.
Perez-Serrano
J
J
Bernadina
W E
WE
Rodriguez-Caabeiro
F
F
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D015153:Blotting, Western; D003429:Cross Reactions; D004591:Electrophoresis, Polyacrylamide Gel; D018840:HSP70 Heat-Shock Proteins / Q000032:analysis* / Q000276:immunology; D007814:Larva / Q000737:chemistry; D016014:Linear Models; D017160:Trichinella spiralis / Q000737:chemistry*
D000911:Antibodies, Monoclonal; D018840:HSP70 Heat-Shock Proteins