{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,3]],"date-time":"2026-05-03T23:44:16Z","timestamp":1777851856671,"version":"3.51.4"},"reference-count":22,"publisher":"SAGE Publications","issue":"10","license":[{"start":{"date-parts":[[2000,10,1]],"date-time":"2000-10-01T00:00:00Z","timestamp":970358400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.sagepub.com\/page\/policies\/text-and-data-mining-license"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["J Histochem Cytochem."],"published-print":{"date-parts":[[2000,10]]},"abstract":"<jats:p>The newly developed Animal Research Kit (ARK) offers a simple and economic way of biotinylating mouse primary antibodies for background-free immunostaining of mouse and rat tissue specimens. Biotinylation involves the use of a biotinylated goat anti-mouse immunoglobulin Fab fragment mixed with a mouse primary antibody and subsequent blocking with normal mouse immunoglobulin. Because a reliable immunoenzyme double staining procedure on human tissue specimens with two unlabeled mouse primary antibodies of identical subclass is almost impossible, we have tested the performance of ARK biotinylation of one primary antibody in a multistep indirect\/direct staining protocol. The multistep double staining procedure involved the subsequent application of an unlabeled mouse monoclonal antibody (MAb) 1 detected with an enzyme-labeled EnVision reagent, normal mouse serum for blocking, followed by a biotinylated mouse MAb 2 and enzyme-labeled streptavidin. Alkaline phosphatase and peroxidase enzymatic activities were developed last. Double staining results obtained with an ARK biotinylated reagent were compared with a truly biotinylated reagent using N-hydroxy succinimide\u2013biotin for conjugation. It appeared that both biotinylation procedures revealed identical double staining results. Although a limited number of antibody combinations have been tested, it is clear that this double staining procedure will be successful for many antibody pairs.<\/jats:p>","DOI":"10.1177\/002215540004801013","type":"journal-article","created":{"date-parts":[[2011,4,1]],"date-time":"2011-04-01T03:17:54Z","timestamp":1301627874000},"page":"1431-1437","source":"Crossref","is-referenced-by-count":20,"title":["The Animal Research Kit (ARK) Can Be Used in a Multistep Double Staining Method for Human Tissue Specimens"],"prefix":"10.1177","volume":"48","author":[{"given":"Chris M. van der","family":"Loos","sequence":"first","affiliation":[{"name":"Academical Medical Centre, Department of Cardiovascular Pathology, Amsterdam, The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Heike","family":"G\u00f6bel","sequence":"additional","affiliation":[{"name":"University of Freiburg, Department of Pathology, Freiburg, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"179","published-online":{"date-parts":[[2000,10,1]]},"reference":[{"key":"bibr1-002215540004801013","first-page":"738","volume":"29","author":"Bisgaard K","year":"1996","journal-title":"Proc 10th Int Congr Histochem Cytochem"},{"key":"bibr2-002215540004801013","volume-title":"Immunohistochemistry 2\u2014A Practical Approach","author":"Brandtzaeg P","year":"1997"},{"key":"bibr3-002215540004801013","doi-asserted-by":"publisher","DOI":"10.1016\/0022-1759(95)00281-2"},{"key":"bibr4-002215540004801013","doi-asserted-by":"publisher","DOI":"10.1177\/38.3.1689333"},{"key":"bibr5-002215540004801013","doi-asserted-by":"publisher","DOI":"10.1177\/35.12.2824601"},{"key":"bibr6-002215540004801013","doi-asserted-by":"publisher","DOI":"10.1007\/978-94-009-3299-9_3"},{"key":"bibr7-002215540004801013","doi-asserted-by":"publisher","DOI":"10.1177\/16.9.557"},{"key":"bibr8-002215540004801013","doi-asserted-by":"publisher","DOI":"10.1177\/37.8.2754250"},{"key":"bibr9-002215540004801013","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2249.1990.tb03309.x"},{"key":"bibr10-002215540004801013","doi-asserted-by":"publisher","DOI":"10.1136\/jcp.51.7.506"},{"key":"bibr11-002215540004801013","doi-asserted-by":"publisher","DOI":"10.1177\/002215549704500301"},{"key":"bibr12-002215540004801013","doi-asserted-by":"publisher","DOI":"10.1177\/27.11.92498"},{"key":"bibr13-002215540004801013","first-page":"457","volume":"45","author":"Tidman N","year":"1981","journal-title":"Clin Exp Immunol"},{"key":"bibr14-002215540004801013","doi-asserted-by":"publisher","DOI":"10.1177\/38.7.2355173"},{"key":"bibr15-002215540004801013","doi-asserted-by":"publisher","DOI":"10.1007\/BF01041348"},{"key":"bibr16-002215540004801013","unstructured":"Van der Loos CM (1998) Immunoenzymatic Double Staining Methods. 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