Open AccessCross‐blot: a rapid screening procedure for determining specificity of antibodies to native proteins of the brown‐rot fungus Postia placenta *
Author(s) -
Clausen Carol A.,
Green Frederick,
Highley Terry L.
Publication year - 1991
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1991.tb04462.x
Subject(s) - polyclonal antibodies , western blot , antibody , antigen , monoclonal antibody , placenta , blot , biology , microbiology and biotechnology , dot blot , biochemistry , immunology , fetus , pregnancy , genetics , gene , dna
Summary A method is described for rapidly screening large numbers of mono‐ or polyclonal antibodies for specificity with an equivalent number of antigens on a single sheet of nitrocellulose paper. This method, referred to as cross‐blot, uses only 120 μl of antigen or antibody and can be completed in less than 6 h. Cross‐blot was developed for rapid screening of hybridoma culture supernatants and can be adapted for use with Western blots of native and denatured proteins as well as screening polyclonal antibodies. In this study, murine monoclonal antibodies were produced to Postia placenta , a brown‐rot wood decay fungus. Immunizing antigens were derived from P. placenta ‐decayed wood extracts. The antibodies were screened for specificity to a number of native proteins produced by the fungus. Cross‐blot proved to an efficient method of visualizing cross‐reacting antibodies while screening large numbers of antibodies for specificity.