{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,30]],"date-time":"2026-05-30T02:35:47Z","timestamp":1780108547075,"version":"3.54.0"},"reference-count":21,"publisher":"Wiley","issue":"6","license":[{"start":{"date-parts":[[2019,3,12]],"date-time":"2019-03-12T00:00:00Z","timestamp":1552348800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/100010926","name":"Illinois State University","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100010926","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Journal of Anatomy"],"published-print":{"date-parts":[[2019,6]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>c\u2010Fos is the product of a gene expressed within neurons in the brain that serves as an anatomical marker of cellular activation. Immunohistochemical staining for c\u2010fos allows a characterization of the effects of many different types of experimental manipulations on neuronal activity, making it a powerful technique for understanding brain, drug and behavior relationships. This study compared visualization of an anti\u2010c\u2010fos primary antibody in 40\u2010\u03bcm\u2010thick cryostat sections of formaldehyde\u2010fixed rat brainstem using either a peroxidase enzyme\u2010conjugated secondary antibody (indirect peroxidase) or the peroxidase\u2010conjugated avidin\u2010biotin complex (<jats:styled-content style=\"fixed-case\">ABC<\/jats:styled-content>) method. All sections were treated with H<jats:sub>2<\/jats:sub>O<jats:sub>2<\/jats:sub> to quench endogenous peroxidase enzyme and sodium borohydride to enhance permeability of the tissue and improve staining quality. Every other section was used to examine either the indirect peroxidase or the <jats:styled-content style=\"fixed-case\">ABC<\/jats:styled-content> method. Sections for the indirect peroxidase method were treated with Triton X\u2010100 detergent to increase tissue permeability, goat serum to reduce non\u2010specific binding of the secondary antibody and, in some cases, bovine serum albumin (<jats:styled-content style=\"fixed-case\">BSA<\/jats:styled-content>) to reduce non\u2010specific binding of the primary antibody. Sections for the <jats:styled-content style=\"fixed-case\">ABC<\/jats:styled-content> method were treated with dilute normal serum, and avidin and biotin solutions and, in some cases <jats:styled-content style=\"fixed-case\">BSA<\/jats:styled-content>. Alternate sections were incubated for 72\u00a0h in either rabbit anti\u2010c\u2010fos primary antibody (1\u00a0:\u00a020\u00a0000) or its vehicle (negative control). For the indirect peroxidase protocol, tissues were treated with peroxidase\u2010conjugated goat anti\u2010rabbit secondary antibody. For the <jats:styled-content style=\"fixed-case\">ABC<\/jats:styled-content> protocol, tissues were treated with biotinylated goat anti\u2010rabbit secondary antibody and <jats:styled-content style=\"fixed-case\">ABC<\/jats:styled-content> peroxidase complex. All sections were reacted with 3,3\u2032\u2010diaminobenzadine (<jats:styled-content style=\"fixed-case\">DAB<\/jats:styled-content>) and H<jats:sub>2<\/jats:sub>O<jats:sub>2<\/jats:sub>, mounted and coverslipped. Both methods produced specific staining of c\u2010fos\u2010containing neurons, relative to the negative control sections. The indirect peroxidase protocol produced clear staining of c\u2010fos\u2010containing neurons, with very little background in the negative control sections. Staining for c\u2010fos was enhanced using the <jats:styled-content style=\"fixed-case\">ABC<\/jats:styled-content> method in that c\u2010fos stained neurons were darker and more clearly visible after shorter treatment with <jats:styled-content style=\"fixed-case\">DAB<\/jats:styled-content>. However, negative control sections showed a greater amount of non\u2010specific staining with the <jats:styled-content style=\"fixed-case\">ABC<\/jats:styled-content> method. Thus, the <jats:styled-content style=\"fixed-case\">ABC<\/jats:styled-content> method was more sensitive but showed reduced specificity, with <jats:styled-content style=\"fixed-case\">BSA<\/jats:styled-content> treatment slightly reducing the level of non\u2010specific staining. Overall, the <jats:styled-content style=\"fixed-case\">ABC<\/jats:styled-content> method produced better visualization and contrast of c\u2010fos\u2010containing neurons against the background color of the tissue.<\/jats:p>","DOI":"10.1111\/joa.12967","type":"journal-article","created":{"date-parts":[[2019,3,12]],"date-time":"2019-03-12T22:11:45Z","timestamp":1552428705000},"page":"936-942","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Comparison of indirect peroxidase and avidin\u2010biotin\u2010peroxidase complex (<scp>ABC<\/scp>) immunohistochemical staining procedures for c\u2010fos in rat brain"],"prefix":"10.1111","volume":"234","author":[{"given":"Jae L.","family":"Butler","sequence":"first","affiliation":[{"name":"Medical Laboratory Science Program Department of Health Sciences Illinois State University Normal IL USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Beverly 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