A novel method for large-scale immuno-SEM using protein G coupled polystyrene beads.
Polystyrene beads with a mean diameter of 0.76 μm were coupled with protein G and then anti-type II collagen IgG or anti-chondroitin-4-sulphate IgG were tagged to protein G. Antibody-tagged beads were applied to articular cartilage and labelled beads were counted in each sample. Antibody-tagged beads labelled significantly higher than IgG isotype control. We propose immuno-SEM using protein G coupled beads as a valuable method for micrometre range observation for specific protein distribution on surfaces of tissues or organs. This will provide information about structure as well as antigenicity on the surface at the same time.
Song
In-Hwan
IH
Department of Anatomy, College of Medicine, Yeungnam University, 317-1 Daemyungdong, Daegu 705-717, South Korea.
Dennis
James E
JE
D000818:Animals; D002358:Cartilage, Articular / Q000378:metabolism*; D002809:Chondroitin Sulfates / Q000276:immunology / Q000378:metabolism; D024043:Collagen Type II / Q000276:immunology / Q000378:metabolism; D005269:Femur; D007118:Immunoassay / Q000379:methods; D007074:Immunoglobulin G / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D008855:Microscopy, Electron, Scanning / Q000379:methods; D009419:Nerve Tissue Proteins / Q000737:chemistry* / Q000235:genetics; D011137:Polystyrenes / Q000737:chemistry*; D011817:Rabbits
D024043:Collagen Type II; C029152:G-substrate; D007074:Immunoglobulin G; D009419:Nerve Tissue Proteins; D011137:Polystyrenes; D002809:Chondroitin Sulfates