Removal of detergents from protein extracts using activated charcoal prior to immunological analysis.
The use of dextran-coated activated charcoal (DCC) powder to absorb solubilising detergents from cell lysates is described. Normal embryonic epithelial cells were lysed in the presence of sodium dodecyl sulphate (SDS). The detergent was then absorbed with DCC to facilitate analysis of polycystin-1 with antibody-based methods. Polycystin-1 is a membrane protein that is involved in the pathogenesis of autosomal dominant polycystic kidney disease (ADPKD). The adverse effect of SDS on antibody-polycystin-1 binding was studied and the improvement resulting from its removal demonstrated using enzyme-linked immunosorbent assays (ELISAs). The results indicate that DCC can be used in a simple manner to remove highly reactive membrane-solubilising reagents from protein mixtures prior to immunological analysis. This procedure may be relevant to a variety of other techniques that are normally affected by detergents.
Division of Life Sciences, King's College London, 150 Stamford Street, London, UK.
Ashraf N
AN
Malhas
Ramadan A
RA
Abuknesha
Robert G
RG
Price
D000327:Adsorption / Q000187:drug effects; D001666:Binding Sites, Antibody; D001667:Binding, Competitive / Q000276:immunology; D002458:Cell Fractionation; D002460:Cell Line; D002606:Charcoal; D002793:Cholic Acids / Q000494:pharmacology; D003902:Detergents / Q000494:pharmacology; D003911:Dextrans; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006801:Humans; D011092:Polyethylene Glycols / Q000494:pharmacology; D011506:Proteins / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification*; D012967:Sodium Dodecyl Sulfate; D050396:TRPP Cation Channels
D002606:Charcoal; D002793:Cholic Acids; D003902:Detergents; D003911:Dextrans; D011092:Polyethylene Glycols; D011506:Proteins; D012967:Sodium Dodecyl Sulfate; D050396:TRPP Cation Channels; C005092:lubrol; C028213:3-((3-cholamidopropyl)dimethylammonium)-1-propanesulfonate