Stabilization of the tetramethylbenzidine (TMB) reaction product: application for retrograde and anterograde tracing, and combination with immunohistochemistry
Tetramethylbenzidine (TMB) as a substrate for horseradish peroxidase (HRP) histochemistry is more sensitive than other chromogens. Its instability in aqueous solutions and ethanol, however, has limited its application. We now report a method for stabilizing TMB by incubation in combinations of diami...
Gespeichert in:
| Veröffentlicht in: | The journal of histochemistry and cytochemistry 1984-11, Vol.32 (11), p.1145-1153 |
|---|---|
| Hauptverfasser: | , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 1153 |
|---|---|
| container_issue | 11 |
| container_start_page | 1145 |
| container_title | The journal of histochemistry and cytochemistry |
| container_volume | 32 |
| creator | Rye, DB Saper, CB Wainer, BH |
| description | Tetramethylbenzidine (TMB) as a substrate for horseradish peroxidase (HRP) histochemistry is more sensitive than other chromogens. Its instability in aqueous solutions and ethanol, however, has limited its application. We now report a method for stabilizing TMB by incubation in combinations of diaminobenzidine (DAB)/cobalt (Co2+)/H2O2. The stabilized TMB product was unaffected by long-term exposures to ethanol, neutral buffers, and subsequent immunohistochemical staining procedures. A procedure is recommended for optimal stabilization of TMB that affords a sensitivity for demonstrating retrogradely labeled perikarya comparable to standard TMB histochemistry. The physical characteristics of the reaction product make it suitable for combination with the unlabeled antibody, peroxidase-antiperoxidase (PAP) immunohistochemical staining procedure. This was established by staining retrogradely labeled neurons in the basal forebrain with a monoclonal antibody against choline acetyltransferase. Because the stabilized TMB product exhibited a superior sensitivity over cobalt ion intensification of the DAB-based reaction product (DAB-Co), it offers a distinct advantage over previously described combination procedures. |
| doi_str_mv | 10.1177/32.11.6548485 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_81315025</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sage_id>10.1177_32.11.6548485</sage_id><sourcerecordid>81315025</sourcerecordid><originalsourceid>FETCH-LOGICAL-c423t-7ab2490b6c51a6dadeac2f69ea6b57d3622c2f85a219f611b1e13c4f917f3a873</originalsourceid><addsrcrecordid>eNp1kctu1TAQhi0EKofCkiVSFoibSPH4kgs7qLhJRSwo62jiOCeuEvtgO4pOX4ZXxTShrFhYv2bmm3_sMSGPgZ4BlOUbzpKeFVJUopJ3yA6khFxSIe6SHaWM5Skh7pMHIVxRCkLI6oScbPiO_PoesTWjucZonM1cn8VBZ1FHj5OOw3Fstb02nbE6e3H59f3LzGtUN-jBu25W8W2Gh8No1NrfO5-I6N3eY6cztF06Uf-Nk6sydv_6pqDc1Bq79i0mDpmZptm6wYTo1KCnpP74kNzrcQz60aan5MfHD5fnn_OLb5--nL-7yJVgPOYltkzUtC2UBCy6NAsV64taY9HKsuMFYymuJDKo-wKgBQ1cib6GsudYlfyUPFt907N-zjrEJs1XehzRajeHpgIOkjKZwHwFlXcheN03B28m9McGaPPnQxrOkjbbhhP_ZDOe20l3t_S_-tOtjkHh2Hu0yoRbrAaQHGjCXq1YwL1urtzsbVrHf2c-X-HB7IfFeN2ECccx3QCaZVlWGEBI_hux3rD7</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>81315025</pqid></control><display><type>article</type><title>Stabilization of the tetramethylbenzidine (TMB) reaction product: application for retrograde and anterograde tracing, and combination with immunohistochemistry</title><source>MEDLINE</source><source>Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals</source><source>SAGE Complete A-Z List</source><source>Free Full-Text Journals in Chemistry</source><creator>Rye, DB ; Saper, CB ; Wainer, BH</creator><creatorcontrib>Rye, DB ; Saper, CB ; Wainer, BH</creatorcontrib><description>Tetramethylbenzidine (TMB) as a substrate for horseradish peroxidase (HRP) histochemistry is more sensitive than other chromogens. Its instability in aqueous solutions and ethanol, however, has limited its application. We now report a method for stabilizing TMB by incubation in combinations of diaminobenzidine (DAB)/cobalt (Co2+)/H2O2. The stabilized TMB product was unaffected by long-term exposures to ethanol, neutral buffers, and subsequent immunohistochemical staining procedures. A procedure is recommended for optimal stabilization of TMB that affords a sensitivity for demonstrating retrogradely labeled perikarya comparable to standard TMB histochemistry. The physical characteristics of the reaction product make it suitable for combination with the unlabeled antibody, peroxidase-antiperoxidase (PAP) immunohistochemical staining procedure. This was established by staining retrogradely labeled neurons in the basal forebrain with a monoclonal antibody against choline acetyltransferase. Because the stabilized TMB product exhibited a superior sensitivity over cobalt ion intensification of the DAB-based reaction product (DAB-Co), it offers a distinct advantage over previously described combination procedures.</description><identifier>ISSN: 0022-1554</identifier><identifier>EISSN: 1551-5044</identifier><identifier>DOI: 10.1177/32.11.6548485</identifier><identifier>PMID: 6548485</identifier><identifier>CODEN: JHCYAS</identifier><language>eng</language><publisher>Los Angeles, CA: Histochemical Soc</publisher><subject>Anatomy ; Animals ; Benzidines ; Biological and medical sciences ; Central nervous system ; Cerebral Cortex - metabolism ; Drug Stability ; Fundamental and applied biological sciences. Psychology ; Histocytochemistry - methods ; Horseradish Peroxidase ; Immunochemistry - methods ; Lectins ; Male ; Rats ; Rats, Inbred Strains ; Temperature ; Vertebrates: nervous system and sense organs ; Wheat Germ Agglutinins</subject><ispartof>The journal of histochemistry and cytochemistry, 1984-11, Vol.32 (11), p.1145-1153</ispartof><rights>1985 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c423t-7ab2490b6c51a6dadeac2f69ea6b57d3622c2f85a219f611b1e13c4f917f3a873</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://journals.sagepub.com/doi/pdf/10.1177/32.11.6548485$$EPDF$$P50$$Gsage$$H</linktopdf><linktohtml>$$Uhttps://journals.sagepub.com/doi/10.1177/32.11.6548485$$EHTML$$P50$$Gsage$$H</linktohtml><link.rule.ids>314,778,782,21806,27911,27912,43608,43609</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=9115310$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/6548485$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Rye, DB</creatorcontrib><creatorcontrib>Saper, CB</creatorcontrib><creatorcontrib>Wainer, BH</creatorcontrib><title>Stabilization of the tetramethylbenzidine (TMB) reaction product: application for retrograde and anterograde tracing, and combination with immunohistochemistry</title><title>The journal of histochemistry and cytochemistry</title><addtitle>J Histochem Cytochem</addtitle><description>Tetramethylbenzidine (TMB) as a substrate for horseradish peroxidase (HRP) histochemistry is more sensitive than other chromogens. Its instability in aqueous solutions and ethanol, however, has limited its application. We now report a method for stabilizing TMB by incubation in combinations of diaminobenzidine (DAB)/cobalt (Co2+)/H2O2. The stabilized TMB product was unaffected by long-term exposures to ethanol, neutral buffers, and subsequent immunohistochemical staining procedures. A procedure is recommended for optimal stabilization of TMB that affords a sensitivity for demonstrating retrogradely labeled perikarya comparable to standard TMB histochemistry. The physical characteristics of the reaction product make it suitable for combination with the unlabeled antibody, peroxidase-antiperoxidase (PAP) immunohistochemical staining procedure. This was established by staining retrogradely labeled neurons in the basal forebrain with a monoclonal antibody against choline acetyltransferase. Because the stabilized TMB product exhibited a superior sensitivity over cobalt ion intensification of the DAB-based reaction product (DAB-Co), it offers a distinct advantage over previously described combination procedures.</description><subject>Anatomy</subject><subject>Animals</subject><subject>Benzidines</subject><subject>Biological and medical sciences</subject><subject>Central nervous system</subject><subject>Cerebral Cortex - metabolism</subject><subject>Drug Stability</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Histocytochemistry - methods</subject><subject>Horseradish Peroxidase</subject><subject>Immunochemistry - methods</subject><subject>Lectins</subject><subject>Male</subject><subject>Rats</subject><subject>Rats, Inbred Strains</subject><subject>Temperature</subject><subject>Vertebrates: nervous system and sense organs</subject><subject>Wheat Germ Agglutinins</subject><issn>0022-1554</issn><issn>1551-5044</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1984</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp1kctu1TAQhi0EKofCkiVSFoibSPH4kgs7qLhJRSwo62jiOCeuEvtgO4pOX4ZXxTShrFhYv2bmm3_sMSGPgZ4BlOUbzpKeFVJUopJ3yA6khFxSIe6SHaWM5Skh7pMHIVxRCkLI6oScbPiO_PoesTWjucZonM1cn8VBZ1FHj5OOw3Fstb02nbE6e3H59f3LzGtUN-jBu25W8W2Gh8No1NrfO5-I6N3eY6cztF06Uf-Nk6sydv_6pqDc1Bq79i0mDpmZptm6wYTo1KCnpP74kNzrcQz60aan5MfHD5fnn_OLb5--nL-7yJVgPOYltkzUtC2UBCy6NAsV64taY9HKsuMFYymuJDKo-wKgBQ1cib6GsudYlfyUPFt907N-zjrEJs1XehzRajeHpgIOkjKZwHwFlXcheN03B28m9McGaPPnQxrOkjbbhhP_ZDOe20l3t_S_-tOtjkHh2Hu0yoRbrAaQHGjCXq1YwL1urtzsbVrHf2c-X-HB7IfFeN2ECccx3QCaZVlWGEBI_hux3rD7</recordid><startdate>198411</startdate><enddate>198411</enddate><creator>Rye, DB</creator><creator>Saper, CB</creator><creator>Wainer, BH</creator><general>Histochemical Soc</general><general>SAGE Publications</general><general>Histochemical Society</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>198411</creationdate><title>Stabilization of the tetramethylbenzidine (TMB) reaction product: application for retrograde and anterograde tracing, and combination with immunohistochemistry</title><author>Rye, DB ; Saper, CB ; Wainer, BH</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c423t-7ab2490b6c51a6dadeac2f69ea6b57d3622c2f85a219f611b1e13c4f917f3a873</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1984</creationdate><topic>Anatomy</topic><topic>Animals</topic><topic>Benzidines</topic><topic>Biological and medical sciences</topic><topic>Central nervous system</topic><topic>Cerebral Cortex - metabolism</topic><topic>Drug Stability</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Histocytochemistry - methods</topic><topic>Horseradish Peroxidase</topic><topic>Immunochemistry - methods</topic><topic>Lectins</topic><topic>Male</topic><topic>Rats</topic><topic>Rats, Inbred Strains</topic><topic>Temperature</topic><topic>Vertebrates: nervous system and sense organs</topic><topic>Wheat Germ Agglutinins</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Rye, DB</creatorcontrib><creatorcontrib>Saper, CB</creatorcontrib><creatorcontrib>Wainer, BH</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>The journal of histochemistry and cytochemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Rye, DB</au><au>Saper, CB</au><au>Wainer, BH</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Stabilization of the tetramethylbenzidine (TMB) reaction product: application for retrograde and anterograde tracing, and combination with immunohistochemistry</atitle><jtitle>The journal of histochemistry and cytochemistry</jtitle><addtitle>J Histochem Cytochem</addtitle><date>1984-11</date><risdate>1984</risdate><volume>32</volume><issue>11</issue><spage>1145</spage><epage>1153</epage><pages>1145-1153</pages><issn>0022-1554</issn><eissn>1551-5044</eissn><coden>JHCYAS</coden><abstract>Tetramethylbenzidine (TMB) as a substrate for horseradish peroxidase (HRP) histochemistry is more sensitive than other chromogens. Its instability in aqueous solutions and ethanol, however, has limited its application. We now report a method for stabilizing TMB by incubation in combinations of diaminobenzidine (DAB)/cobalt (Co2+)/H2O2. The stabilized TMB product was unaffected by long-term exposures to ethanol, neutral buffers, and subsequent immunohistochemical staining procedures. A procedure is recommended for optimal stabilization of TMB that affords a sensitivity for demonstrating retrogradely labeled perikarya comparable to standard TMB histochemistry. The physical characteristics of the reaction product make it suitable for combination with the unlabeled antibody, peroxidase-antiperoxidase (PAP) immunohistochemical staining procedure. This was established by staining retrogradely labeled neurons in the basal forebrain with a monoclonal antibody against choline acetyltransferase. Because the stabilized TMB product exhibited a superior sensitivity over cobalt ion intensification of the DAB-based reaction product (DAB-Co), it offers a distinct advantage over previously described combination procedures.</abstract><cop>Los Angeles, CA</cop><pub>Histochemical Soc</pub><pmid>6548485</pmid><doi>10.1177/32.11.6548485</doi><tpages>9</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0022-1554 |
| ispartof | The journal of histochemistry and cytochemistry, 1984-11, Vol.32 (11), p.1145-1153 |
| issn | 0022-1554 1551-5044 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_81315025 |
| source | MEDLINE; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; SAGE Complete A-Z List; Free Full-Text Journals in Chemistry |
| subjects | Anatomy Animals Benzidines Biological and medical sciences Central nervous system Cerebral Cortex - metabolism Drug Stability Fundamental and applied biological sciences. Psychology Histocytochemistry - methods Horseradish Peroxidase Immunochemistry - methods Lectins Male Rats Rats, Inbred Strains Temperature Vertebrates: nervous system and sense organs Wheat Germ Agglutinins |
| title | Stabilization of the tetramethylbenzidine (TMB) reaction product: application for retrograde and anterograde tracing, and combination with immunohistochemistry |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-15T20%3A36%3A37IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Stabilization%20of%20the%20tetramethylbenzidine%20(TMB)%20reaction%20product:%20application%20for%20retrograde%20and%20anterograde%20tracing,%20and%20combination%20with%20immunohistochemistry&rft.jtitle=The%20journal%20of%20histochemistry%20and%20cytochemistry&rft.au=Rye,%20DB&rft.date=1984-11&rft.volume=32&rft.issue=11&rft.spage=1145&rft.epage=1153&rft.pages=1145-1153&rft.issn=0022-1554&rft.eissn=1551-5044&rft.coden=JHCYAS&rft_id=info:doi/10.1177/32.11.6548485&rft_dat=%3Cproquest_cross%3E81315025%3C/proquest_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=81315025&rft_id=info:pmid/6548485&rft_sage_id=10.1177_32.11.6548485&rfr_iscdi=true |