Characterization of Rat Gastric Mucins Using a Monoclonal Antibody, RGM23, Recognizing Surface Mucous Cell–Type Mucins
A novel anti-mucin monoclonal antibody (mAb), designated RGM23, was developed against mucin purified from rat gastric mucosa. RGM23 reacted with the mucin attached to the ELISA well. The reactivity was lost by trypsin treatment, but not by periodate oxidation, indicating that RGM23 recognizes the pe...
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| Veröffentlicht in: | Journal of biochemistry (Tokyo) 2003-04, Vol.133 (4), p.453-460 |
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| container_title | Journal of biochemistry (Tokyo) |
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| creator | Goso, Yukinobu Ikezawa, Tomoaki Kurihara, Makoto Endo, Masaki Hotta, Kyoko Ishihara, Kazuhiko |
| description | A novel anti-mucin monoclonal antibody (mAb), designated RGM23, was developed against mucin purified from rat gastric mucosa. RGM23 reacted with the mucin attached to the ELISA well. The reactivity was lost by trypsin treatment, but not by periodate oxidation, indicating that RGM23 recognizes the peptide moiety of the mucin molecule. Histochemical study showed that RGM23 stained the corpus and antral surface mucosa of rat stomach, but not their glandular mucosa, nor duodenal, small intestinal or large intestinal mucosa. The area stained with RGM23 was coincident with that stained with 45M1, a mAb reacting with MUC5AC mucin. Examination of the mucin subunits extracted from rat stomach by Sepharose CL-4B and Q-Sepharose chromatography and CsTFA equilibrium centrifugation showed that RGM23 reacted with the surface mucous cell-type mucins that were stained with periodate-Schiff (PAS) and reacted with mAb RGM21. The gastric gland-type mucin, which reacted with mAb HIK1083, did not react with RGM23. On Q-Sepharose chromatography, a part of the RGM21-reactive mucins was only faintly stained with PAS and did not react with RGM23. The results together indicated that RGM23 probably reacted with the rat MUC5AC (rMuc5AC) mucin present in the surface mucosa of the stomach, and that the surface mucosal cells in rat stomach may contain mucin bearing non-rMuc5AC core protein in addition to rMuc5AC mucins. |
| doi_str_mv | 10.1093/jb/mvg059 |
| format | Article |
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RGM23 reacted with the mucin attached to the ELISA well. The reactivity was lost by trypsin treatment, but not by periodate oxidation, indicating that RGM23 recognizes the peptide moiety of the mucin molecule. Histochemical study showed that RGM23 stained the corpus and antral surface mucosa of rat stomach, but not their glandular mucosa, nor duodenal, small intestinal or large intestinal mucosa. The area stained with RGM23 was coincident with that stained with 45M1, a mAb reacting with MUC5AC mucin. Examination of the mucin subunits extracted from rat stomach by Sepharose CL-4B and Q-Sepharose chromatography and CsTFA equilibrium centrifugation showed that RGM23 reacted with the surface mucous cell-type mucins that were stained with periodate-Schiff (PAS) and reacted with mAb RGM21. The gastric gland-type mucin, which reacted with mAb HIK1083, did not react with RGM23. On Q-Sepharose chromatography, a part of the RGM21-reactive mucins was only faintly stained with PAS and did not react with RGM23. The results together indicated that RGM23 probably reacted with the rat MUC5AC (rMuc5AC) mucin present in the surface mucosa of the stomach, and that the surface mucosal cells in rat stomach may contain mucin bearing non-rMuc5AC core protein in addition to rMuc5AC mucins.</description><identifier>ISSN: 0021-924X</identifier><identifier>DOI: 10.1093/jb/mvg059</identifier><identifier>PMID: 12761292</identifier><language>eng</language><publisher>England: Oxford University Press</publisher><subject>Animals ; Antibodies, Monoclonal - immunology ; Antibody Specificity ; Centrifugation, Density Gradient ; cesium trifluoroacetate ; Chromatography, Agarose - methods ; CsTFA ; dithiothreitol ; DTT ; Gastric Mucins - chemistry ; Gastric Mucins - immunology ; Gastric Mucosa - chemistry ; Gastric Mucosa - immunology ; glandular mucosa ; guanidinium hydrochloride ; GuHCl ; horseradish peroxidase ; HRP ; Immunohistochemistry ; Key words: gastric mucin ; mAb ; monoclonal antibody ; Mucin 5AC ; Mucins - analysis ; PAS ; periodate-Schiff ; phenylmethylsulphonyl fluoride ; PMSF ; rat MUC5AC mucin ; Rats ; rMuc5AC ; Stomach - chemistry ; Stomach - immunology ; surface mucosa ; Trypsin</subject><ispartof>Journal of biochemistry (Tokyo), 2003-04, Vol.133 (4), p.453-460</ispartof><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c368t-747e664e2cef9290e014aef51973272d39de9e5829fa8bf4c10705736417b8ff3</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,27901,27902</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/12761292$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Goso, Yukinobu</creatorcontrib><creatorcontrib>Ikezawa, Tomoaki</creatorcontrib><creatorcontrib>Kurihara, Makoto</creatorcontrib><creatorcontrib>Endo, Masaki</creatorcontrib><creatorcontrib>Hotta, Kyoko</creatorcontrib><creatorcontrib>Ishihara, Kazuhiko</creatorcontrib><title>Characterization of Rat Gastric Mucins Using a Monoclonal Antibody, RGM23, Recognizing Surface Mucous Cell–Type Mucins</title><title>Journal of biochemistry (Tokyo)</title><addtitle>J Biochem</addtitle><description>A novel anti-mucin monoclonal antibody (mAb), designated RGM23, was developed against mucin purified from rat gastric mucosa. RGM23 reacted with the mucin attached to the ELISA well. The reactivity was lost by trypsin treatment, but not by periodate oxidation, indicating that RGM23 recognizes the peptide moiety of the mucin molecule. Histochemical study showed that RGM23 stained the corpus and antral surface mucosa of rat stomach, but not their glandular mucosa, nor duodenal, small intestinal or large intestinal mucosa. The area stained with RGM23 was coincident with that stained with 45M1, a mAb reacting with MUC5AC mucin. Examination of the mucin subunits extracted from rat stomach by Sepharose CL-4B and Q-Sepharose chromatography and CsTFA equilibrium centrifugation showed that RGM23 reacted with the surface mucous cell-type mucins that were stained with periodate-Schiff (PAS) and reacted with mAb RGM21. The gastric gland-type mucin, which reacted with mAb HIK1083, did not react with RGM23. On Q-Sepharose chromatography, a part of the RGM21-reactive mucins was only faintly stained with PAS and did not react with RGM23. The results together indicated that RGM23 probably reacted with the rat MUC5AC (rMuc5AC) mucin present in the surface mucosa of the stomach, and that the surface mucosal cells in rat stomach may contain mucin bearing non-rMuc5AC core protein in addition to rMuc5AC mucins.</description><subject>Animals</subject><subject>Antibodies, Monoclonal - immunology</subject><subject>Antibody Specificity</subject><subject>Centrifugation, Density Gradient</subject><subject>cesium trifluoroacetate</subject><subject>Chromatography, Agarose - methods</subject><subject>CsTFA</subject><subject>dithiothreitol</subject><subject>DTT</subject><subject>Gastric Mucins - chemistry</subject><subject>Gastric Mucins - immunology</subject><subject>Gastric Mucosa - chemistry</subject><subject>Gastric Mucosa - immunology</subject><subject>glandular mucosa</subject><subject>guanidinium hydrochloride</subject><subject>GuHCl</subject><subject>horseradish peroxidase</subject><subject>HRP</subject><subject>Immunohistochemistry</subject><subject>Key words: gastric mucin</subject><subject>mAb</subject><subject>monoclonal antibody</subject><subject>Mucin 5AC</subject><subject>Mucins - analysis</subject><subject>PAS</subject><subject>periodate-Schiff</subject><subject>phenylmethylsulphonyl fluoride</subject><subject>PMSF</subject><subject>rat MUC5AC mucin</subject><subject>Rats</subject><subject>rMuc5AC</subject><subject>Stomach - chemistry</subject><subject>Stomach - immunology</subject><subject>surface mucosa</subject><subject>Trypsin</subject><issn>0021-924X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2003</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpFkM1O4zAUhb0ADT_DghdAXiEhkal_kjheogpaUKuROkVCbCzHvS4uaVzsBLWseAfecJ5kUrViVkdX9ztHRwehc0p-USJ5b1H2lu9zkskDdEwIo4lk6dMROolxsT0Z5z_QEWUip0yyY7Tuv-igTQPBfejG-Rp7iye6wQMdm-AMHrfG1RE_RlfPscZjX3tT-VpX-KZuXOlnm2s8GYwZ7wSMn9fuY0v-aYPVBrZ230bch6r6-_k13axgn_gTHVpdRTjb6yl6vLud9ofJ6Pfgvn8zSgzPiyYRqYA8T4EZsJJJAoSmGmxGpeBMsBmXM5CQFUxaXZQ2NZQIkgmep1SUhbX8FF3uclfBv7UQG7V00XR1dA1dMyU4pwXL8w682oEm-BgDWLUKbqnDRlGittOqRal203bsxT60LZcw-0_ud-2AZAe42MD6-6_Dq8oFF5kaPj0rOR2OhmJM1QP_B_wkhys</recordid><startdate>200304</startdate><enddate>200304</enddate><creator>Goso, Yukinobu</creator><creator>Ikezawa, Tomoaki</creator><creator>Kurihara, Makoto</creator><creator>Endo, Masaki</creator><creator>Hotta, Kyoko</creator><creator>Ishihara, Kazuhiko</creator><general>Oxford University Press</general><scope>BSCLL</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>200304</creationdate><title>Characterization of Rat Gastric Mucins Using a Monoclonal Antibody, RGM23, Recognizing Surface Mucous Cell–Type Mucins</title><author>Goso, Yukinobu ; Ikezawa, Tomoaki ; Kurihara, Makoto ; Endo, Masaki ; Hotta, Kyoko ; Ishihara, Kazuhiko</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c368t-747e664e2cef9290e014aef51973272d39de9e5829fa8bf4c10705736417b8ff3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2003</creationdate><topic>Animals</topic><topic>Antibodies, Monoclonal - immunology</topic><topic>Antibody Specificity</topic><topic>Centrifugation, Density Gradient</topic><topic>cesium trifluoroacetate</topic><topic>Chromatography, Agarose - methods</topic><topic>CsTFA</topic><topic>dithiothreitol</topic><topic>DTT</topic><topic>Gastric Mucins - chemistry</topic><topic>Gastric Mucins - immunology</topic><topic>Gastric Mucosa - chemistry</topic><topic>Gastric Mucosa - immunology</topic><topic>glandular mucosa</topic><topic>guanidinium hydrochloride</topic><topic>GuHCl</topic><topic>horseradish peroxidase</topic><topic>HRP</topic><topic>Immunohistochemistry</topic><topic>Key words: gastric mucin</topic><topic>mAb</topic><topic>monoclonal antibody</topic><topic>Mucin 5AC</topic><topic>Mucins - analysis</topic><topic>PAS</topic><topic>periodate-Schiff</topic><topic>phenylmethylsulphonyl fluoride</topic><topic>PMSF</topic><topic>rat MUC5AC mucin</topic><topic>Rats</topic><topic>rMuc5AC</topic><topic>Stomach - chemistry</topic><topic>Stomach - immunology</topic><topic>surface mucosa</topic><topic>Trypsin</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Goso, Yukinobu</creatorcontrib><creatorcontrib>Ikezawa, Tomoaki</creatorcontrib><creatorcontrib>Kurihara, Makoto</creatorcontrib><creatorcontrib>Endo, Masaki</creatorcontrib><creatorcontrib>Hotta, Kyoko</creatorcontrib><creatorcontrib>Ishihara, Kazuhiko</creatorcontrib><collection>Istex</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>Journal of biochemistry (Tokyo)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Goso, Yukinobu</au><au>Ikezawa, Tomoaki</au><au>Kurihara, Makoto</au><au>Endo, Masaki</au><au>Hotta, Kyoko</au><au>Ishihara, Kazuhiko</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Characterization of Rat Gastric Mucins Using a Monoclonal Antibody, RGM23, Recognizing Surface Mucous Cell–Type Mucins</atitle><jtitle>Journal of biochemistry (Tokyo)</jtitle><addtitle>J Biochem</addtitle><date>2003-04</date><risdate>2003</risdate><volume>133</volume><issue>4</issue><spage>453</spage><epage>460</epage><pages>453-460</pages><issn>0021-924X</issn><abstract>A novel anti-mucin monoclonal antibody (mAb), designated RGM23, was developed against mucin purified from rat gastric mucosa. RGM23 reacted with the mucin attached to the ELISA well. The reactivity was lost by trypsin treatment, but not by periodate oxidation, indicating that RGM23 recognizes the peptide moiety of the mucin molecule. Histochemical study showed that RGM23 stained the corpus and antral surface mucosa of rat stomach, but not their glandular mucosa, nor duodenal, small intestinal or large intestinal mucosa. The area stained with RGM23 was coincident with that stained with 45M1, a mAb reacting with MUC5AC mucin. Examination of the mucin subunits extracted from rat stomach by Sepharose CL-4B and Q-Sepharose chromatography and CsTFA equilibrium centrifugation showed that RGM23 reacted with the surface mucous cell-type mucins that were stained with periodate-Schiff (PAS) and reacted with mAb RGM21. The gastric gland-type mucin, which reacted with mAb HIK1083, did not react with RGM23. On Q-Sepharose chromatography, a part of the RGM21-reactive mucins was only faintly stained with PAS and did not react with RGM23. The results together indicated that RGM23 probably reacted with the rat MUC5AC (rMuc5AC) mucin present in the surface mucosa of the stomach, and that the surface mucosal cells in rat stomach may contain mucin bearing non-rMuc5AC core protein in addition to rMuc5AC mucins.</abstract><cop>England</cop><pub>Oxford University Press</pub><pmid>12761292</pmid><doi>10.1093/jb/mvg059</doi><tpages>8</tpages></addata></record> |
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| source | J-STAGE Free; Oxford University Press Journals All Titles (1996-Current); MEDLINE; Free Full-Text Journals in Chemistry |
| subjects | Animals Antibodies, Monoclonal - immunology Antibody Specificity Centrifugation, Density Gradient cesium trifluoroacetate Chromatography, Agarose - methods CsTFA dithiothreitol DTT Gastric Mucins - chemistry Gastric Mucins - immunology Gastric Mucosa - chemistry Gastric Mucosa - immunology glandular mucosa guanidinium hydrochloride GuHCl horseradish peroxidase HRP Immunohistochemistry Key words: gastric mucin mAb monoclonal antibody Mucin 5AC Mucins - analysis PAS periodate-Schiff phenylmethylsulphonyl fluoride PMSF rat MUC5AC mucin Rats rMuc5AC Stomach - chemistry Stomach - immunology surface mucosa Trypsin |
| title | Characterization of Rat Gastric Mucins Using a Monoclonal Antibody, RGM23, Recognizing Surface Mucous Cell–Type Mucins |
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