{"id":16,"date":"2015-07-27T01:10:40","date_gmt":"2015-07-27T06:10:40","guid":{"rendered":"http:\/\/www.creative-diagnostics.com\/blog\/?p=16"},"modified":"2020-12-11T00:58:02","modified_gmt":"2020-12-11T05:58:02","slug":"elisa-a-vital-technology-in-modern-experiment1","status":"publish","type":"post","link":"https:\/\/www.creative-diagnostics.com\/blog\/index.php\/elisa-a-vital-technology-in-modern-experiment1\/","title":{"rendered":"ELISA &#8211;A Vital Technology in Modern Experiment(1)"},"content":{"rendered":"<h2>Relatively\u00a0New\u00a0Methods\u00a0of\u00a0ELISA<\/h2>\n<p style=\"text-align: justify;\">ELISA, created in 60s-70s in last century, has got a wide good reputation from researchers for its stable reagents, easy operation, objective diagnosis. ELISA biological test is an accurate, sensible, specific testing method. Thus application area of ELISA is expanding rapidly.<\/p>\n<h3>Theory<\/h3>\n<p style=\"text-align: justify;\">Immunoassay is based on special binding affinities produced by antibodies and antigens, thus any high-quality reagents won\u2019t be produced without good materials such as antibody and enzyme. In the past, the antigens which were applied to immunoassay usually were various purified antigens, and the antibodies of the assay were monoclonal antibody produced by purifying antigen immune animals as well as polyclonal antibody produced by hybridoma technique. Recently, new materials are appearing. For example, some specific antigens or antibodies are made by gene engineering and new immunoassay reagents which are produced by mixture of enzyme. There are various new methods including double-antibody sandwich and indirect assay.<\/p>\n<h3>Double-antibody Sandwich Assay<\/h3>\n<p style=\"text-align: justify;\">Double-antibody Sandwich Assay, a test for antigen or an application of the<a href=\"\/ELISA-Kits.htm\"> ELISA<\/a> method, is usually used in testing antigens of macromolecules. The double-antibody assay generally include the following four steps:<br \/>\nFirst step: absorb antiserum containing known antibody at the ori<img decoding=\"async\" loading=\"lazy\" class=\" size-full wp-image-28 alignright\" src=\"http:\/\/www.creative-diagnostics.com\/blog\/wp-content\/uploads\/2015\/07\/ELISA-sandwich_meitu_21.jpg\" alt=\"ELISA sandwich_meitu_2\" width=\"270\" height=\"238\" \/>fices of microtiter plate and wash up extra antibody.<br \/>\nSecond step:add testing antigen. If the two are special,they are bind. Then wash up the extra antibody.<br \/>\nThird step:add adding antibody (secondary antibody) linked to the enzyme with special reaction and make it sandwich.<br \/>\nFourth step: add substrate of enzyme to check whether the colored enzymatic hydrolysate is produced. If it is, which means that the antigen exist at walls of orifices.<\/p>\n<h3><strong>Indirect Assay<\/strong><\/h3>\n<p>The indirect method of <a href=\"\/ELISA-Kits.htm\">ELISA<\/a> is thus better suited for diagnostic work. There are generally following four steps:<br \/>\nFirst step, coat known antigen with microtiter plate and wash extra antigen.<br \/>\nSecond step, add sample to check whether there is primary antibody w<img decoding=\"async\" loading=\"lazy\" class=\" wp-image-27 alignright\" src=\"http:\/\/www.creative-diagnostics.com\/blog\/wp-content\/uploads\/2015\/07\/indirect-ELISA-_meitu_5.jpg\" alt=\"indirect ELISA _meitu_5\" width=\"347\" height=\"259\" \/>hich is to be tested. If it is, the sample will have specific binding. Then wash up extra sample.<br \/>\nThird step:add secondary antibody linked with enzymes to have specific binding with primary antibody which is to be tested.<br \/>\nFourth step, wash up extra secondary antibody without specific binding and add enzymes to make enzymes colored. Then test Optical Density in microtiter plate by ELISA reader to estimate the antibody\u2019s content which is to be tested in ultimate product.<\/p>\n<p><a href=\"\/blog\/index.php\/elisa-a-vital-technology-in-modern-experiment-2\/\">To be continued<\/a><\/p>\n","protected":false},"excerpt":{"rendered":"<p>Relatively\u00a0New\u00a0Methods\u00a0of\u00a0ELISA ELISA, created in 60s-70s in last century, has got a wide good reputation from researchers for its stable reagents, easy operation, objective diagnosis. ELISA biological test is an accurate, sensible, specific testing method. Thus application area of ELISA is expanding rapidly. Theory Immunoassay is based on special binding affinities produced by antibodies and antigens, [&hellip;]<\/p>\n","protected":false},"author":1,"featured_media":0,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":[],"categories":[59],"tags":[73],"aioseo_notices":[],"_links":{"self":[{"href":"https:\/\/www.creative-diagnostics.com\/blog\/index.php\/wp-json\/wp\/v2\/posts\/16"}],"collection":[{"href":"https:\/\/www.creative-diagnostics.com\/blog\/index.php\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/www.creative-diagnostics.com\/blog\/index.php\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/www.creative-diagnostics.com\/blog\/index.php\/wp-json\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/www.creative-diagnostics.com\/blog\/index.php\/wp-json\/wp\/v2\/comments?post=16"}],"version-history":[{"count":7,"href":"https:\/\/www.creative-diagnostics.com\/blog\/index.php\/wp-json\/wp\/v2\/posts\/16\/revisions"}],"predecessor-version":[{"id":960,"href":"https:\/\/www.creative-diagnostics.com\/blog\/index.php\/wp-json\/wp\/v2\/posts\/16\/revisions\/960"}],"wp:attachment":[{"href":"https:\/\/www.creative-diagnostics.com\/blog\/index.php\/wp-json\/wp\/v2\/media?parent=16"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.creative-diagnostics.com\/blog\/index.php\/wp-json\/wp\/v2\/categories?post=16"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.creative-diagnostics.com\/blog\/index.php\/wp-json\/wp\/v2\/tags?post=16"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}