title: A sensitive and reproducible method for the detection of oligoclonal IgM bands.
abstract: We have developed a sensitive and specific isoelectrofocusing (IEF) method for the detection of oligoclonal (OGC) IgM in the cerebrospinal fluid (CSF) and sera. In this procedure, ampholytes, in the range pH 5-8, were used to improve the resolution, and serum was diluted in saline to avoid IgM precipitation. Samples were treated with 50 mM dithiothreitol (DTT) at pH 9.5 to reduce IgM and improve the migration of the protein. Using an anti-IgM labelled with alkaline phosphatase, the limit of detection was found to be 0.1 ng in 5 microl of sample.
authors.affiliation: Department of Immunology, Hospital Ramón y Cajal, Madrid, Spain. lvillar@hrc.insalud.es
authors.forename: L M
authors.initials: LM
authors.lastname: Villar
authors.forename: P
authors.initials: P
authors.lastname: González-Porqué
authors.forename: J
authors.initials: J
authors.lastname: Masjuán
authors.forename: J C
authors.initials: JC
authors.lastname: Alvarez-Cermeño
authors.forename: A
authors.initials: A
authors.lastname: Bootello
authors.forename: G
authors.initials: G
authors.lastname: Keir
mesh_terms: D002021:Buffers; D004229:Dithiothreitol; D006801:Humans; D006863:Hydrogen-Ion Concentration; D015151:Immunoblotting; D007075:Immunoglobulin M / Q000032:analysis* / Q000097:blood / Q000134:cerebrospinal fluid; D007136:Immunoglobulins / Q000032:analysis* / Q000097:blood / Q000134:cerebrospinal fluid; D007525:Isoelectric Focusing / Q000379:methods* / Q000706:statistics & numerical data; D009103:Multiple Sclerosis / Q000276:immunology; D043025:Oligoclonal Bands; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D002021:Buffers; D007075:Immunoglobulin M; D007136:Immunoglobulins; D043025:Oligoclonal Bands; D004229:Dithiothreitol