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filingDate 1991-02-27-04:00^^<http://www.w3.org/2001/XMLSchema#date>
grantDate 1994-06-21-04:00^^<http://www.w3.org/2001/XMLSchema#date>
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publicationDate 1994-06-21-04:00^^<http://www.w3.org/2001/XMLSchema#date>
publicationNumber US-5322771-A
titleOfInvention Immunohistochemical staining method and reagents therefor
abstract PCT No. PCT/US91/01108 Sec. 371 Date Aug. 31, 1992 Sec. 102(e) Date Aug. 31, 1992 PCT Filed Feb. 27, 1991 PCT Pub. No. WO91/13336 PCT Pub. Date Sep. 5, 1991.The present invention provides an improved method for staining slides using immunochemical reagents. The method comprises the following steps. The assay region of a slide (the region containing the tissue section) is washed with an improved rinsing solution comprising water and a detergent. An evaporation inhibitor liquid is applied to the slide to cover the assay region. For antigens requiring unmasking, the tissue section is combined with an improved, stabilized proteolytic enzyme solution. The slide is rinsed, and the evaporation inhibitor liquid is reapplied to the slide. A primary antibody in an improved diluent containing globulins from the same species as a second antibody is combined with the tissue section for a time sufficient for substantially complete antibody binding. The slide is rinsed, and the evaporation inhibitor liquid is reapplied. A labeled second antibody in the improved diluent is combined with the tissue section for a time sufficient for substantially complete antibody binding. The slide is rinsed, and the evaporation inhibitor liquid is reapplied to the slide. Color development reagents, including, in a preferred embodiment, a stabilized diaminobenzidine (DAB) solution, are combined with the tissue section for a time sufficient for color development. Following rinsing, the tissue section is ready for analysis.
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