Preparation and use of recombinant protein G-gold complexes as markers in double labelling immunocytochemistry
Recombinant protein G (RPG) was conjugated to colloidal gold particles and used for immunocytochemistry. In this report, the preparation of RPG-gold conjugates (RPGG) and the application of these conjugates in spot blot tests and in double immunolabelling are described. The immunolabelling was perfo...
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| Veröffentlicht in: | The Histochemical journal 1989-08, Vol.21 (8), p.449-454 |
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| creator | BALSLEV, Y HANSEN, G. H |
| description | Recombinant protein G (RPG) was conjugated to colloidal gold particles and used for immunocytochemistry. In this report, the preparation of RPG-gold conjugates (RPGG) and the application of these conjugates in spot blot tests and in double immunolabelling are described. The immunolabelling was performed on ultracryosections of pig small intestine using antibodies directed against aminopeptidase N and sucrase-isomaltase. The labelling efficiency of RPGG was compared to that of protein A-gold conjugates (PAG) in different compartments of the enterocyte. Quantification showed that the labelling intensity was dependent on the size of the marker as well as on the kind of protein used for complex formation. The distributions for RPGG and PAG were respectively: for the 12 nm particles, 10.3 and 6.2 particles/micron of length of microvillar membrane, 3.5 and 1.0 particles/micron2 of Golgi profile and 5.9 and 2.0 particles/micron2 of multivesicular body profile; and for the 6 nm particles, 49.6 and 15.7 particles/micron of length of microvillar membrane, 24.4 and 5.0 particles/micron2 of Golgi profile and 25.4 and 3.4 particles/micron2 of multivesicular body profile. Controls showed very little non-specific gold labelling (less than 0.02 gold particles/micron2 of section). |
| doi_str_mv | 10.1007/BF01845794 |
| format | Article |
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The distributions for RPGG and PAG were respectively: for the 12 nm particles, 10.3 and 6.2 particles/micron of length of microvillar membrane, 3.5 and 1.0 particles/micron2 of Golgi profile and 5.9 and 2.0 particles/micron2 of multivesicular body profile; and for the 6 nm particles, 49.6 and 15.7 particles/micron of length of microvillar membrane, 24.4 and 5.0 particles/micron2 of Golgi profile and 25.4 and 3.4 particles/micron2 of multivesicular body profile. 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H</creatorcontrib><title>Preparation and use of recombinant protein G-gold complexes as markers in double labelling immunocytochemistry</title><title>The Histochemical journal</title><addtitle>Histochem J</addtitle><description>Recombinant protein G (RPG) was conjugated to colloidal gold particles and used for immunocytochemistry. In this report, the preparation of RPG-gold conjugates (RPGG) and the application of these conjugates in spot blot tests and in double immunolabelling are described. The immunolabelling was performed on ultracryosections of pig small intestine using antibodies directed against aminopeptidase N and sucrase-isomaltase. The labelling efficiency of RPGG was compared to that of protein A-gold conjugates (PAG) in different compartments of the enterocyte. Quantification showed that the labelling intensity was dependent on the size of the marker as well as on the kind of protein used for complex formation. The distributions for RPGG and PAG were respectively: for the 12 nm particles, 10.3 and 6.2 particles/micron of length of microvillar membrane, 3.5 and 1.0 particles/micron2 of Golgi profile and 5.9 and 2.0 particles/micron2 of multivesicular body profile; and for the 6 nm particles, 49.6 and 15.7 particles/micron of length of microvillar membrane, 24.4 and 5.0 particles/micron2 of Golgi profile and 25.4 and 3.4 particles/micron2 of multivesicular body profile. Controls showed very little non-specific gold labelling (less than 0.02 gold particles/micron2 of section).</description><subject>Analytical, structural and metabolic biochemistry</subject><subject>Animals</subject><subject>Bacterial Proteins - metabolism</subject><subject>Binding and carrier proteins</subject><subject>Biological and medical sciences</subject><subject>Biomarkers</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gold - metabolism</subject><subject>Immunohistochemistry - methods</subject><subject>Intestinal Mucosa - metabolism</subject><subject>Intestines - cytology</subject><subject>Proteins</subject><subject>Recombinant Proteins - metabolism</subject><subject>Swine</subject><issn>0018-2214</issn><issn>1573-6865</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1989</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpFkM1LAzEQxYMotVYv3oUcxIOwmuxHkj1q0SoU9KDnZTY7qau7SU12wf73RlrqXAbm_Zh58wg55-yGMyZv7x8ZV3khy_yATHkhs0QoURySKYvzJE15fkxOQvhkjJVSigmZpIrJUrEpsa8e1-BhaJ2lYBs6BqTOUI_a9XVrwQ507d2AraWLZOW6hkZh3eEPBgqB9uC_0Aca5caNdYe0gxq7rrUr2vb9aJ3eDE5_YN-GwW9OyZGBLuDZrs_I--PD2_wpWb4snud3y0RnnA8JQIrAoc4MprkWWBrdgDAyZ0qLJh6QmSqAca0ykxZMSCZKjVinNatNrmU2I1fbvdH794hhqOJ9HX2BRTeGSpapKgQvIni9BbV3IXg01dq38alNxVn1F271H26EL3Zbx7rHZo_u0oz65U6HoKEzHqxuwx4TZa5iZb-pnoPd</recordid><startdate>19890801</startdate><enddate>19890801</enddate><creator>BALSLEV, Y</creator><creator>HANSEN, G. 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H</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c311t-aa2ea1ab3fe24c6e9fcda6f7408c6dabe7385a01c83f25067069ceeb2b0bf4c73</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1989</creationdate><topic>Analytical, structural and metabolic biochemistry</topic><topic>Animals</topic><topic>Bacterial Proteins - metabolism</topic><topic>Binding and carrier proteins</topic><topic>Biological and medical sciences</topic><topic>Biomarkers</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Gold - metabolism</topic><topic>Immunohistochemistry - methods</topic><topic>Intestinal Mucosa - metabolism</topic><topic>Intestines - cytology</topic><topic>Proteins</topic><topic>Recombinant Proteins - metabolism</topic><topic>Swine</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>BALSLEV, Y</creatorcontrib><creatorcontrib>HANSEN, G. H</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 Histochemical journal</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>BALSLEV, Y</au><au>HANSEN, G. H</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Preparation and use of recombinant protein G-gold complexes as markers in double labelling immunocytochemistry</atitle><jtitle>The Histochemical journal</jtitle><addtitle>Histochem J</addtitle><date>1989-08-01</date><risdate>1989</risdate><volume>21</volume><issue>8</issue><spage>449</spage><epage>454</epage><pages>449-454</pages><issn>0018-2214</issn><eissn>1573-6865</eissn><coden>HISJAE</coden><abstract>Recombinant protein G (RPG) was conjugated to colloidal gold particles and used for immunocytochemistry. In this report, the preparation of RPG-gold conjugates (RPGG) and the application of these conjugates in spot blot tests and in double immunolabelling are described. The immunolabelling was performed on ultracryosections of pig small intestine using antibodies directed against aminopeptidase N and sucrase-isomaltase. The labelling efficiency of RPGG was compared to that of protein A-gold conjugates (PAG) in different compartments of the enterocyte. Quantification showed that the labelling intensity was dependent on the size of the marker as well as on the kind of protein used for complex formation. The distributions for RPGG and PAG were respectively: for the 12 nm particles, 10.3 and 6.2 particles/micron of length of microvillar membrane, 3.5 and 1.0 particles/micron2 of Golgi profile and 5.9 and 2.0 particles/micron2 of multivesicular body profile; and for the 6 nm particles, 49.6 and 15.7 particles/micron of length of microvillar membrane, 24.4 and 5.0 particles/micron2 of Golgi profile and 25.4 and 3.4 particles/micron2 of multivesicular body profile. Controls showed very little non-specific gold labelling (less than 0.02 gold particles/micron2 of section).</abstract><cop>London</cop><pub>Kluwer</pub><pmid>2807980</pmid><doi>10.1007/BF01845794</doi><tpages>6</tpages></addata></record> |
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| identifier | ISSN: 0018-2214 |
| ispartof | The Histochemical journal, 1989-08, Vol.21 (8), p.449-454 |
| issn | 0018-2214 1573-6865 |
| language | eng |
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| source | MEDLINE; SpringerLink Journals - AutoHoldings |
| subjects | Analytical, structural and metabolic biochemistry Animals Bacterial Proteins - metabolism Binding and carrier proteins Biological and medical sciences Biomarkers Fundamental and applied biological sciences. Psychology Gold - metabolism Immunohistochemistry - methods Intestinal Mucosa - metabolism Intestines - cytology Proteins Recombinant Proteins - metabolism Swine |
| title | Preparation and use of recombinant protein G-gold complexes as markers in double labelling immunocytochemistry |
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