A microtiter-based assay for the detection of protein tyrosine kinase activity
A 96-well microtiter enzyme-linked immunosorbent assay (ELISA) for protein tyrosine kinases has been developed. This assay uses one of several substrates that are phosphorylated by tyrosine kinase, an antibody to phosphotyrosine, and a peroxidase-linked second antibody. Color development is monitore...
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| Veröffentlicht in: | Analytical biochemistry 1990-11, Vol.190 (2), p.249-253 |
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| creator | Cleaveland, Jeffrey S. Kiener, Peter A. Hammond, David J. Schacter, Bernice Z. |
| description | A 96-well microtiter enzyme-linked immunosorbent assay (ELISA) for protein tyrosine kinases has been developed. This assay uses one of several substrates that are phosphorylated by tyrosine kinase, an antibody to phosphotyrosine, and a peroxidase-linked second antibody. Color development is monitored by a change in absorbance at 450 nm and is dependent upon time, enzyme, ATP, and substrate concentrations. Specificity of the ELISA for phosphotyrosine was shown by inhibition of binding of the anti-phosphotyrosine antibody with phenyl phosphate. Results obtained in the ELISA compared favorably with those obtained by direct phosphorylation of the substrate with [
32P]ATP. Staurosporine and K252a, protein kinase inhibitors, showed titratable inhibition of tyrosine kinase activity. This assay is a rapid, nonradioactive alternative to traditional methodology and is also amenable to automation. |
| doi_str_mv | 10.1016/0003-2697(90)90188-F |
| format | Article |
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32P]ATP. Staurosporine and K252a, protein kinase inhibitors, showed titratable inhibition of tyrosine kinase activity. This assay is a rapid, nonradioactive alternative to traditional methodology and is also amenable to automation.</description><identifier>ISSN: 0003-2697</identifier><identifier>EISSN: 1096-0309</identifier><identifier>DOI: 10.1016/0003-2697(90)90188-F</identifier><identifier>PMID: 1705396</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>550201 - Biochemistry- Tracer Techniques ; Adenosine Triphosphate - metabolism ; Alkaloids - pharmacology ; AMINO ACIDS ; Antibodies - immunology ; Autoanalysis ; BASIC BIOLOGICAL SCIENCES ; BETA DECAY RADIOISOTOPES ; BETA-MINUS DECAY RADIOISOTOPES ; Binding, Competitive ; BIOASSAY ; Carbazoles - pharmacology ; CARBOXYLIC ACIDS ; CHEMICAL REACTIONS ; DAYS LIVING RADIOISOTOPES ; ENZYME ACTIVITY ; ENZYME IMMUNOASSAY ; ENZYME INHIBITORS ; Enzyme Stability - drug effects ; Enzyme-Linked Immunosorbent Assay ; ENZYMES ; ErbB Receptors - physiology ; HYDROXY ACIDS ; IMMUNOASSAY ; Indole Alkaloids ; ISOTOPE APPLICATIONS ; ISOTOPES ; LIGHT NUCLEI ; Muramidase - metabolism ; NUCLEI ; ODD-ODD NUCLEI ; ORGANIC ACIDS ; ORGANIC COMPOUNDS ; Phosphoric Monoester Hydrolases - metabolism ; PHOSPHORUS 32 ; PHOSPHORUS ISOTOPES ; Phosphorus Radioisotopes ; PHOSPHORUS-GROUP TRANSFERASES ; PHOSPHORYLATION ; PHOSPHOTRANSFERASES ; Phosphotyrosine ; Protein-Tyrosine Kinases - analysis ; Protein-Tyrosine Kinases - immunology ; RADIOISOTOPES ; Staurosporine ; Substrate Specificity - drug effects ; TRACER TECHNIQUES ; TRANSFERASES ; TYROSINE ; Tyrosine - analogs & derivatives ; Tyrosine - immunology</subject><ispartof>Analytical biochemistry, 1990-11, Vol.190 (2), p.249-253</ispartof><rights>1990</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c384t-c90a81fe1dd54cc6edd9afa4317d77c868e7d74f9a8dd844280cd67af50ed673</citedby><cites>FETCH-LOGICAL-c384t-c90a81fe1dd54cc6edd9afa4317d77c868e7d74f9a8dd844280cd67af50ed673</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/0003-2697(90)90188-F$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>230,315,781,785,886,3551,27929,27930,46000</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/1705396$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink><backlink>$$Uhttps://www.osti.gov/biblio/5962728$$D View this record in Osti.gov$$Hfree_for_read</backlink></links><search><creatorcontrib>Cleaveland, Jeffrey S.</creatorcontrib><creatorcontrib>Kiener, Peter A.</creatorcontrib><creatorcontrib>Hammond, David J.</creatorcontrib><creatorcontrib>Schacter, Bernice Z.</creatorcontrib><title>A microtiter-based assay for the detection of protein tyrosine kinase activity</title><title>Analytical biochemistry</title><addtitle>Anal Biochem</addtitle><description>A 96-well microtiter enzyme-linked immunosorbent assay (ELISA) for protein tyrosine kinases has been developed. This assay uses one of several substrates that are phosphorylated by tyrosine kinase, an antibody to phosphotyrosine, and a peroxidase-linked second antibody. Color development is monitored by a change in absorbance at 450 nm and is dependent upon time, enzyme, ATP, and substrate concentrations. Specificity of the ELISA for phosphotyrosine was shown by inhibition of binding of the anti-phosphotyrosine antibody with phenyl phosphate. Results obtained in the ELISA compared favorably with those obtained by direct phosphorylation of the substrate with [
32P]ATP. Staurosporine and K252a, protein kinase inhibitors, showed titratable inhibition of tyrosine kinase activity. This assay is a rapid, nonradioactive alternative to traditional methodology and is also amenable to automation.</description><subject>550201 - Biochemistry- Tracer Techniques</subject><subject>Adenosine Triphosphate - metabolism</subject><subject>Alkaloids - pharmacology</subject><subject>AMINO ACIDS</subject><subject>Antibodies - immunology</subject><subject>Autoanalysis</subject><subject>BASIC BIOLOGICAL SCIENCES</subject><subject>BETA DECAY RADIOISOTOPES</subject><subject>BETA-MINUS DECAY RADIOISOTOPES</subject><subject>Binding, Competitive</subject><subject>BIOASSAY</subject><subject>Carbazoles - pharmacology</subject><subject>CARBOXYLIC ACIDS</subject><subject>CHEMICAL REACTIONS</subject><subject>DAYS LIVING RADIOISOTOPES</subject><subject>ENZYME ACTIVITY</subject><subject>ENZYME IMMUNOASSAY</subject><subject>ENZYME INHIBITORS</subject><subject>Enzyme Stability - drug effects</subject><subject>Enzyme-Linked Immunosorbent Assay</subject><subject>ENZYMES</subject><subject>ErbB Receptors - physiology</subject><subject>HYDROXY ACIDS</subject><subject>IMMUNOASSAY</subject><subject>Indole Alkaloids</subject><subject>ISOTOPE APPLICATIONS</subject><subject>ISOTOPES</subject><subject>LIGHT NUCLEI</subject><subject>Muramidase - metabolism</subject><subject>NUCLEI</subject><subject>ODD-ODD NUCLEI</subject><subject>ORGANIC ACIDS</subject><subject>ORGANIC COMPOUNDS</subject><subject>Phosphoric Monoester Hydrolases - metabolism</subject><subject>PHOSPHORUS 32</subject><subject>PHOSPHORUS ISOTOPES</subject><subject>Phosphorus Radioisotopes</subject><subject>PHOSPHORUS-GROUP TRANSFERASES</subject><subject>PHOSPHORYLATION</subject><subject>PHOSPHOTRANSFERASES</subject><subject>Phosphotyrosine</subject><subject>Protein-Tyrosine Kinases - analysis</subject><subject>Protein-Tyrosine Kinases - immunology</subject><subject>RADIOISOTOPES</subject><subject>Staurosporine</subject><subject>Substrate Specificity - drug effects</subject><subject>TRACER TECHNIQUES</subject><subject>TRANSFERASES</subject><subject>TYROSINE</subject><subject>Tyrosine - analogs & derivatives</subject><subject>Tyrosine - immunology</subject><issn>0003-2697</issn><issn>1096-0309</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1990</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kE1LxDAQhoMoun78A4XgQfRQnWzbNLkIsrgqiF68h5hMMbrbrElW6L83tYvePE1gnnfy8hByzOCSAeNXAFAWUy6bcwkXEpgQxXyLTBhIXkAJcptMfpE9sh_jOwBjVc13yS5roC4ln5CnG7p0JvjkEobiVUe0VMeoe9r6QNMbUosJTXK-o76lq0yi62jqg4-uQ_rhupyhOhNfLvWHZKfVi4hHm3lAXua3L7P74vH57mF281iYUlSpMBK0YC0ya-vKGI7WSt3qqmSNbRojuMD8qFqphbWiqqYCjOWNbmvAPMsDcjqe9TE5FU0ub96M77rcVNWST5upyNDZCOXSn2uMSS1dNLhY6A79OioB2UzJ6gxWI5g9xBiwVavgljr0ioEaVKvBoxo8KgnqR7Wa59jJ5v76dYn2LzS6zfvrcY9ZxJfDMBTFzqB1Yehpvfv_g2_en449</recordid><startdate>19901101</startdate><enddate>19901101</enddate><creator>Cleaveland, Jeffrey S.</creator><creator>Kiener, Peter A.</creator><creator>Hammond, David J.</creator><creator>Schacter, Bernice Z.</creator><general>Elsevier Inc</general><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><scope>OTOTI</scope></search><sort><creationdate>19901101</creationdate><title>A microtiter-based assay for the detection of protein tyrosine kinase activity</title><author>Cleaveland, Jeffrey S. ; Kiener, Peter A. ; Hammond, David J. ; Schacter, Bernice Z.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c384t-c90a81fe1dd54cc6edd9afa4317d77c868e7d74f9a8dd844280cd67af50ed673</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1990</creationdate><topic>550201 - Biochemistry- Tracer Techniques</topic><topic>Adenosine Triphosphate - metabolism</topic><topic>Alkaloids - pharmacology</topic><topic>AMINO ACIDS</topic><topic>Antibodies - immunology</topic><topic>Autoanalysis</topic><topic>BASIC BIOLOGICAL SCIENCES</topic><topic>BETA DECAY RADIOISOTOPES</topic><topic>BETA-MINUS DECAY RADIOISOTOPES</topic><topic>Binding, Competitive</topic><topic>BIOASSAY</topic><topic>Carbazoles - pharmacology</topic><topic>CARBOXYLIC ACIDS</topic><topic>CHEMICAL REACTIONS</topic><topic>DAYS LIVING RADIOISOTOPES</topic><topic>ENZYME ACTIVITY</topic><topic>ENZYME IMMUNOASSAY</topic><topic>ENZYME INHIBITORS</topic><topic>Enzyme Stability - drug effects</topic><topic>Enzyme-Linked Immunosorbent Assay</topic><topic>ENZYMES</topic><topic>ErbB Receptors - physiology</topic><topic>HYDROXY ACIDS</topic><topic>IMMUNOASSAY</topic><topic>Indole Alkaloids</topic><topic>ISOTOPE APPLICATIONS</topic><topic>ISOTOPES</topic><topic>LIGHT NUCLEI</topic><topic>Muramidase - metabolism</topic><topic>NUCLEI</topic><topic>ODD-ODD NUCLEI</topic><topic>ORGANIC ACIDS</topic><topic>ORGANIC COMPOUNDS</topic><topic>Phosphoric Monoester Hydrolases - metabolism</topic><topic>PHOSPHORUS 32</topic><topic>PHOSPHORUS ISOTOPES</topic><topic>Phosphorus Radioisotopes</topic><topic>PHOSPHORUS-GROUP TRANSFERASES</topic><topic>PHOSPHORYLATION</topic><topic>PHOSPHOTRANSFERASES</topic><topic>Phosphotyrosine</topic><topic>Protein-Tyrosine Kinases - analysis</topic><topic>Protein-Tyrosine Kinases - immunology</topic><topic>RADIOISOTOPES</topic><topic>Staurosporine</topic><topic>Substrate Specificity - drug effects</topic><topic>TRACER TECHNIQUES</topic><topic>TRANSFERASES</topic><topic>TYROSINE</topic><topic>Tyrosine - analogs & derivatives</topic><topic>Tyrosine - immunology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Cleaveland, Jeffrey S.</creatorcontrib><creatorcontrib>Kiener, Peter A.</creatorcontrib><creatorcontrib>Hammond, David J.</creatorcontrib><creatorcontrib>Schacter, Bernice Z.</creatorcontrib><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><collection>OSTI.GOV</collection><jtitle>Analytical biochemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Cleaveland, Jeffrey S.</au><au>Kiener, Peter A.</au><au>Hammond, David J.</au><au>Schacter, Bernice Z.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A microtiter-based assay for the detection of protein tyrosine kinase activity</atitle><jtitle>Analytical biochemistry</jtitle><addtitle>Anal Biochem</addtitle><date>1990-11-01</date><risdate>1990</risdate><volume>190</volume><issue>2</issue><spage>249</spage><epage>253</epage><pages>249-253</pages><issn>0003-2697</issn><eissn>1096-0309</eissn><abstract>A 96-well microtiter enzyme-linked immunosorbent assay (ELISA) for protein tyrosine kinases has been developed. This assay uses one of several substrates that are phosphorylated by tyrosine kinase, an antibody to phosphotyrosine, and a peroxidase-linked second antibody. Color development is monitored by a change in absorbance at 450 nm and is dependent upon time, enzyme, ATP, and substrate concentrations. Specificity of the ELISA for phosphotyrosine was shown by inhibition of binding of the anti-phosphotyrosine antibody with phenyl phosphate. Results obtained in the ELISA compared favorably with those obtained by direct phosphorylation of the substrate with [
32P]ATP. Staurosporine and K252a, protein kinase inhibitors, showed titratable inhibition of tyrosine kinase activity. This assay is a rapid, nonradioactive alternative to traditional methodology and is also amenable to automation.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>1705396</pmid><doi>10.1016/0003-2697(90)90188-F</doi><tpages>5</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0003-2697 |
| ispartof | Analytical biochemistry, 1990-11, Vol.190 (2), p.249-253 |
| issn | 0003-2697 1096-0309 |
| language | eng |
| recordid | cdi_osti_scitechconnect_5962728 |
| source | MEDLINE; Access via ScienceDirect (Elsevier) |
| subjects | 550201 - Biochemistry- Tracer Techniques Adenosine Triphosphate - metabolism Alkaloids - pharmacology AMINO ACIDS Antibodies - immunology Autoanalysis BASIC BIOLOGICAL SCIENCES BETA DECAY RADIOISOTOPES BETA-MINUS DECAY RADIOISOTOPES Binding, Competitive BIOASSAY Carbazoles - pharmacology CARBOXYLIC ACIDS CHEMICAL REACTIONS DAYS LIVING RADIOISOTOPES ENZYME ACTIVITY ENZYME IMMUNOASSAY ENZYME INHIBITORS Enzyme Stability - drug effects Enzyme-Linked Immunosorbent Assay ENZYMES ErbB Receptors - physiology HYDROXY ACIDS IMMUNOASSAY Indole Alkaloids ISOTOPE APPLICATIONS ISOTOPES LIGHT NUCLEI Muramidase - metabolism NUCLEI ODD-ODD NUCLEI ORGANIC ACIDS ORGANIC COMPOUNDS Phosphoric Monoester Hydrolases - metabolism PHOSPHORUS 32 PHOSPHORUS ISOTOPES Phosphorus Radioisotopes PHOSPHORUS-GROUP TRANSFERASES PHOSPHORYLATION PHOSPHOTRANSFERASES Phosphotyrosine Protein-Tyrosine Kinases - analysis Protein-Tyrosine Kinases - immunology RADIOISOTOPES Staurosporine Substrate Specificity - drug effects TRACER TECHNIQUES TRANSFERASES TYROSINE Tyrosine - analogs & derivatives Tyrosine - immunology |
| title | A microtiter-based assay for the detection of protein tyrosine kinase activity |
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