title: The application of glucose oxidase-labeled antibodies for the detection of proteins on nitrocellulose.
abstract: We have evaluated the sensitivity of immunostaining with glucose oxidase for the detection of monomeric human serum albumin (HSA) and monomeric human immunoglobulin G (IgG). A modification of a histochemical procedure was utilized by replacing phenazine methosulfate (PMS) with 1-methoxyphenazine methosulfate (mPMS) and by replacing Tris-HCl with Tris-citrate to improve the solubility of the tetrazolium compounds tested. mPMS is less sensitive to light, and may be stored for long periods in solution; it is now used routinely by histochemists in place of PMS in dehydrogenase cytochemistry. pH values of 6.3-8.3 were tested, with the reaction at pH 8.3 providing a slight increase in sensitivity. The reaction rate increased markedly as the pH became more alkaline. The minimum quantity of HSA detected was 3 ng applied directly to nitrocellulose and 10 ng when blotted. Human IgG was routinely detected at 250 pg and occasionally at 100 pg when dotted on the nitrocellulose.
mesh_terms: D003101:Collodion; D005949:Glucose Oxidase; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007124:Immunoenzyme Techniques; D007074:Immunoglobulin G / Q000032:analysis*; D007525:Isoelectric Focusing; D012709:Serum Albumin / Q000032:analysis*
chemical_list: D007074:Immunoglobulin G; D012709:Serum Albumin; D003101:Collodion; D005949:Glucose Oxidase