CINXE.COM

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id="block-block-26" class="block block-block"> <div class="content"> <div class="container_12"><a href="https://www.bio-techne.com/account/novus-bio-techne/login?_ga=2.17257593.151271452<br />
5.1724696046-473375462.1712005244" style="margin: 0 10px;" target="_blank"><img alt="Novus Biologicals products are now on bio-techne.com" src="https://images.novusbio.com/design/ready-to-buy-novus-products-login.png" style="height:120px; width:940px" /></a></div> </div> </div> </div> <h1>Secondary Antibodies – Explore Types of Antibodies</h1> </div> <div class="clear"></div> <div class="grid_16"> <div class="region region-content"> <div id="block-et-messages-et-messages-block" class="block block-et-messages"> <div class="content"> <span></span> </div> </div> <div id="block-system-main" class="block block-system"> <div class="content"> <div id="node-12347146" class="node node-page clearfix"> <div class="content"> <div class="field field-name-body field-type-text-with-summary field-label-hidden"><div class="field-items"><div class="field-item even"><head> <style> .container { position: relative; text-align: center; color: black; } .line2Back { position: absolute; top: 80px; left: 310px; height:19px; width:310px; background-color: #C5CFDF; opacity: 0.5; } .line4Back { position: absolute; top: 160px; left: 310px; height:60px; width:310px; background-color: #C5CFDF; opacity: 0.5; } .line6Back { position: absolute; top: 280px; left: 310px; height:19px; width:310px; background-color: #C5CFDF; opacity: 0.5; } .line1 { position: absolute; top: 60px; left: 320px; } .line1b { position: absolute; top: 60px; left: 440px; } .line2 { position: absolute; top: 80px; left: 320px; z-index: 1; } .line2b { position: absolute; top: 80px; left: 440px; z-index: 1; } .line3 { position: absolute; top: 100px; left: 320px; } .line3b { position: absolute; top: 100px; left: 440px; } .line3c { position: absolute; top: 120px; left: 440px; } .line3d { position: absolute; top: 140px; left: 440px; } .line4 { position: absolute; top: 160px; left: 320px; z-index: 1; } .line4b { position: absolute; top: 160px; left: 440px; z-index: 1; } .line4c { position: absolute; top: 180px; left: 440px; z-index: 1; } .line4d { position: absolute; top: 200px; left: 440px; z-index: 1; } .line5 { position: absolute; top: 220px; left: 320px; } .line5b { position: absolute; top: 220px; left: 440px; } .line5c { position: absolute; top: 240px; left: 440px; } .line5d { position: absolute; top: 260px; left: 440px; } .line6 { position: absolute; top: 280px; left: 320px; z-index: 1; } .line6b { position: absolute; top: 280px; left: 440px; z-index: 1; } .line7 { position: absolute; top: 300px; left: 320px; } .line7b { position: absolute; top: 300px; left: 440px; } .line7c { position: absolute; top: 320px; left: 440px; } .line7d { position: absolute; top: 340px; left: 440px; } </style> </head> <table cellspacing="1" cellpadding="1" width="100%" align="center" name="parent" border="0"> <tr> <td valign="top" width="200px"> <b><u>Related Links</u></b> <p><a href="/product-type/secondary-antibodies">Secondary Antibodies</a></p> <p><a href="/search?facet_conjugate=Janelia%20Fluor%20549&category=Secondary%20Antibodies">Janelia Fluor<sup>®</sup> 549 Conjugated Antibodies</a></p> <p><a href="/search?facet_conjugate=Janelia%20Fluor%20646&category=Secondary%20Antibodies">Janelia Fluor<sup>®</sup> 646 Conjugated Antibodies</a></p> <p><a href="/search?keywords=novus&category=Secondary%20Antibodies&facet_conjugate=HRP">HRP Conjugated Antibodies</a></p> <b><u>Antibody Resources</u></b> <p><a href="/support/support-by-application.html">Protocols</a></p> <p><a href="/support/secondary-antibody-handbook">Secondary Antibody Handbook</a></p> <p><a href="/support/secondary-antibodies-faqs">Secondary Antibody FAQs</a></p> <p><a href="/support/detection-methods-in-icc-if">Direct vs. Indirect Detection</a></p> <p><a href="/products/janelia-fluor">Janelia Fluor<sup>®</sup> Dyes</a></p> </td> <td> <table border="0" width="100%" align="left" cellspacing="8" cellpadding="8"> <tr> <td valign="top" colspan="4"> <tr> <table border="0" cellpadding="4" cellspacing="4"> <p style="margin-left:24px;"> <h2>What is a Secondary Antibody?</h2> <p>A secondary antibody is an antibody directed against another immunoglobulin (antibody) molecule. Under ideal conditions, a secondary antibody specifically binds the species and immunoglobulin (Ig) class (isotype) of the primary antibody in an immunoassay (i.e. indirect detection). The label, or conjugate, of the secondary antibody determines the method of detection, either chromogenic or fluorescent. In addition to detection of primary antibodies, secondaries are also used for antibody capture in ELISAs and for detection of recombinant protein.</p> </td> </tr> <tr> <td colspan="2" valign="top"> <p style="margin-left:24px;"> </td> </tr> <tr> <td colspan="4" valign="top" bgcolor="#c5cfdf"> <p><b>Find your antibody of interest using the search tool below.</b></p> <form action="/search" method="get"> <div class="et_searchbar"><input autocomplete="off" name="keywords" type="text" class="et_searchbar_wide et_searchbar_default" value="" placeholder="" data-default=""><input type="submit" value="search" class="fp_mag_glass_wideSearch"></div> </form> <br/> <p align="right"><a href="/product-type/secondary-antibodies">Browse all secondary antibodies</a></p> </td> </tr> <tr> <table border="0" cellpadding="4" cellspacing="4"> <p>Our secondary antibodies are validated and optimized for use in <a href="/application/western-blotting">western blot (WB)</a>, <a href="/application/icc">immunocytochemistry (ICC)</a>, <a href="/application/ihc">immunohistochemistry (IHC)</a>, <a href="/application/flow-cytometry">flow cytometry</a>, <a href="/application/elisa"> ELISA</a>, and more. </p> </td> </tr> <tr> <td valign="top" width="20%">        <b>Host Species</b> </td> <td valign="top" width="20%">        <b>Reactivity</b> </td> <td valign="top" width="20%">        <b>Target Ig Class</b> </td> <td valign="top" width="20%">        <b>Conjugates</b> </td> </tr> <tr> <td valign="top"> <ul style="list-style-image: url('https://images.novusbio.com/design/psBullet.gif'); margin-top:-6px;"> <li><a href="/product-type/secondary-antibodies?_=1460577397713&fq%5bhost%5d=Goat">Goat</a></li> <li><a href="/product-type/secondary-antibodies?_=1460577397715&fq%5bhost%5d=Sheep">Sheep</a></li> <li><a href="/product-type/secondary-antibodies?_=1460577397717&fq%5bhost%5d=Donkey">Donkey</a></li> <li><a href="/product-type/secondary-antibodies?_=1460577397719&fq%5bhost%5d=Rabbit">Rabbit</a></li> <li><a href="/product-type/secondary-antibodies?_=1460577397721&fq%5bhost%5d=Chicken">Chicken</a></li> <li><a href="/product-type/secondary-antibodies?_=1460577397723&fq%5bhost%5d=Mouse">Mouse</a></li> </ul> </td> <td valign="top"> <ul style="list-style-image: url('https://images.novusbio.com/design/psBullet.gif'); margin-top:-6px;"> <li><a href="/product-type/secondary-antibodies?fq%5bspecies%5d%5b0%5d=Human">Anti-human</a></li> <li><a href="/product-type/secondary-antibodies?_=1460577397705&fq%5bspecies%5d%5b1%5d=Rabbit">Anti-rabbit</a></li> <li><a href="/product-type/secondary-antibodies?_=1460577397703&fq%5bspecies%5d%5b0%5d=Mouse">Anti-mouse</a></li> <li><a href="/product-type/secondary-antibodies?_=1460577397707&fq%5bspecies%5d%5b0%5d=Rat">Anti-rat</a></li> <li><a href="/product-type/secondary-antibodies?_=1460577397709&fq%5bspecies%5d%5b0%5d=Goat">Anti-goat</a></li> <li><a href="/product-type/secondary-antibodies?_=1460577397711&fq%5bspecies%5d%5b0%5d=Sheep">Anti-sheep</a></li> </ul> </td> <td valign="top"> <ul style="list-style-image: url('https://images.novusbio.com/design/psBullet.gif'); margin-top:-6px;"> <li><a href="/product-type/secondary-antibodies?common_name=IgG%20(H%2BL)">IgG</a></li> <li><a href="/product-type/secondary-antibodies?common_name=IgD">IgD</a></li> <li><a href="/product-type/secondary-antibodies?common_name=IgM">IgM</a></li> <li><a href="/product-type/secondary-antibodies?common_name=IgA">IgA</a></li> <li><a href="/product-type/secondary-antibodies?common_name=IgE">IgE</a></li> </ul> </td> <td valign="top"> <ul style="list-style-image: url('https://images.novusbio.com/design/psBullet.gif'); margin-top:-6px;"> <li><a href="/product-type/secondary-antibodies?_=1460577397747&fq%5bconjugate%5d=HRP">HRP</a></li> <li><a href="/product-type/secondary-antibodies?_=1460577397749&fq%5bconjugate%5d=Alkaline%2BPhosphatase">AP</a></li> <li><a href="/product-type/secondary-antibodies?_=1460577397751&fq%5bconjugate%5d=Biotin">Biotin</a></li> <li><a href="/product-type/secondary-antibodies?facet_conjugate=DyLight%20350">DyLight 350</a></li> <li><a href="/product-type/secondary-antibodies?facet_conjugate=DyLight%20488">DyLight 488</a></li> <li><a href="/product-type/secondary-antibodies?facet_conjugate=DyLight%20550">DyLight 550</a></li> <li><a href="/product-type/secondary-antibodies?facet_conjugate=DyLight%20650">DyLight 650</a></li> <li><a href="/product-type/secondary-antibodies?facet_conjugate=DyLight%20800">DyLight 800</a></li> <li><a href="/product-type/secondary-antibodies?_=1460577397741&fq%5bconjugate%5d=FITC">FITC</a></li> <li><a href="/product-type/secondary-antibodies?_=1460577397743&fq%5bconjugate%5d=PE">PE</a></li> </ul> </td> </tr> </table> <br/> <table border="0" cellpadding="4" cellspacing="4"> <tr> <td valign="top" colspan="2"> <p style="margin-left:24px;"> <h2>Secondary Antibody Production</h2> </p> <p style="margin-left:24px;"> Secondary antibody production involves harvesting from an animal that has been immunized with an antibody from another species. The specifications of the immunizing antibody (e.g. species, subclass, fragment, etc.) determine the specificity of the secondary antibody produced. For example, a <a href="/search?keywords=igg%20(h%20l)&category=Secondary%20Antibodies&common_name=IgG%20(H%2BL)&host=Goat&species=Rabbit">goat anti-rabbit IgG (H+L) secondary antibody</a> is produced by immunizing a goat with the whole IgG molecule or heavy and light chains of a rabbit IgG immunoglobulin molecule.</p> </td> </tr> <tr> <td valign="top" colspan="2"> <p style="margin-left:24px;"> <h2>Why Use a Secondary Antibody?</h2> </p> <p style="margin-left:24px;"> Using a secondary antibody increases assay sensitivity and signal amplification due to the ability of multiple secondary antibodies to bind to a single primary antibody. The use of a secondary antibody also provides more flexibility in multiplexing (i.e. multiple labeling) experiments because commercially available secondary antibodies can be pre-conjugated to a wide range of labels.</p> </td> </tr> <tr> <td valign="top" colspan="2"> <p style="margin-left:24px;"> <h2>Antibody Selection: Finding the Right Secondary</h2> </p> <p style="margin-left:24px;">Factors to consider in secondary antibody selection: <ul> <li><strong>Host Species</strong> – the species used to raise the secondary antibody</li> <li><strong>Format</strong> – the structure and fragment of the secondary antibody</li> <li><strong>Species Reactivity</strong> – the species recognized by the secondary antibody</li> <li><strong>Target Immunoglobulin</strong> – The primary antibody class/isotype or subclass</li> <li><strong>Specificity</strong> – The region of the target immunoglobulin recognized by the secondary antibody</li> <li><strong>Conjugate</strong> – the detection label attached to the secondary antibody</li> </ul> </p> <p style="margin-left:24px;">If you cannot find the product you’re looking for Novus Biologicals can help you find the right antibody in addition to providing custom services, such as custom conjugation, to meet your experimental needs.</p> <p align="center"><a class="link_button" "text-center" href="https://www.novusbio.com/support/bulk-and-custom.html">Explore Bulk and Custom Services</a></p> </td> </tr> <tr> <tr> <td colspan="2" valign="top"> <h2>Example of Secondary Antibody Nomenclature</h2> </td> </tr> <tr> <td valign="top" colspan="2"> <div class="container"> <img src="https://images.novusbio.com/design/secondaryAntibodyNomenclature06.png" alt="Goat F(ab) anti-Rabbit lgG1 (H+L) Secondary Antibody [HRP] (Pre-absorbed)" title="Example of Secondary Antibody Nomenclature" style="z-index: -1;"> <div class="line1"><b>Goat:</b></div> <div class="line1b">Host Species</div> <div class="line2" style=""><b>F(ab):</b></div> <div class="line2b">Antibody Structure</div> <div class="line3"><b>Rabbit</b>:</div> <div class="line3b">Species Reactivity</div> <div class="line3c"><i>(Host of the Primary</i></div> <div class="line3d"><i>Antibody)</i></div> <div class="line4"><b>IgG1:</b></div> <div class="line4b">Target Immunoglobulin</div> <div class="line4c"><i>(Class and subclass of</i></div> <div class="line4d"><i>the Primary Antibody)</i></div> <div class="line5"><b>H+L:</b></div> <div class="line5b">Heavy + Light chain,</div> <div class="line5c">Target Specificity <i>(Region</i></div> <div class="line5d"><i>of the Primary Antibody)</i></div> <div class="line6"><b>HRP:</b></div> <div class="line6b">Detection Label</div> <div class="line7"><b>Pre-adsorbed:</b></div> <div class="line7b">Additional Purification</div> <div class="line7c">Step to Reduce Species</div> <div class="line7d">Cross-reactivity</div> <div class="line2Back"></div> <div class="line4Back"></div> <div class="line6Back"></div> </div> <p align="center"><a class="link_button" "text-center" href="https://www.novusbio.com/support/secondary-antibodies-faqs">Explore Secondary Antibodies FAQs</a></p> </td> </tr> <tr> <td valign="top" colspan="2"> <p style="margin-left:24px;"> <h2>Why Choose a Pre-adsorbed Secondary Antibody?</h2> </p> <p style="margin-left:24px;"> Pre-adsorption is a technique used to increase secondary antibody specificity and reduce non-specific binding. The pre-adsorbtion procedure works by passing antibodies through a column containing immobilized serum proteins or immunoglobulin from potentially cross-reactive species. The reactive antibodies bind to the proteins and immunoglobulins, while the non-reactive antibodies flow through the column.<br><br> Pre-adsorbed secondary antibodies are recommended when determining protein expression in multiplexing experiments or when staining immunoglobulin-rich tissues or cells in order to help prevent non-specific binding. It is advised to use a secondary antibody pre-adsorbed against the species of the experimental sample. For example, a secondary antibody adsorbed against rat immunoglobulin or serum is recommended when staining rat tissue.</p> </td> </tr> <tr> <td valign="top" colspan="2"> <p style="margin-left:24px;"> <h2>Janelia Fluor<sup>®</sup> Conjugated Secondary Antibodies</h2> </p> <div class="container"> <img src="//images.novusbio.com/design/janeliaFluorCSABs02.jpg" alt="Janelia Fluor® 549 and Janelia Fluor® 646 conjugated secondary antibodies" title="Janelia Fluor® Conjugated Secondary Antibodies" style="z-index: -1;"> </div> </td> </tr> <tr> <td colspan="2" valign="top"> <p style="margin-left:24px;"> <a href="https://www.novusbio.com/search?facet_conjugate=Janelia%20Fluor%20549&category=Secondary%20Antibodies">Janelia Fluor<sup>®</sup> 549</a> and <a href="https://www.novusbio.com/search?facet_conjugate=Janelia%20Fluor%20646&category=Secondary%20Antibodies">Janelia Fluor<sup>®</sup> 646</a> conjugated secondary antibodies expand the palette of bright, photostable dyes available for your super resolution microscopy techniques including dSTORM and STED imaging experiments. </p> <p align="center"><a class="link_button" "text-center" href="/products/janelia-fluor">Learn More About Janelia Fluor<sup>®</sup> Dyes</a></p> </td> </tr> </table> <br/><br/><br/> </td> </tr> </table></div></div></div> </div> </div> </div> </div> <div id="block-block-21" class="block block-block"> <div class="content">  <div class="embeddedServiceInvitation" id="snapins_invite" aria-live="assertive" role="dialog" aria-atomic="true" style="display: none;"> <div class="embeddedServiceInvitationHeader" aria-labelledby="snapins_titletext" aria-describedby="snapins_bodytext"> <img id="embeddedServiceAvatar" /> <span class="embeddedServiceTitleText" id="snapins_titletext">Need help?</span> <button type="button" id="closeInvite" class="embeddedServiceCloseIcon" aria-label="Exit invitation">×</button> </div> <div class="embeddedServiceInvitationBody"> <p id="snapins_bodytext">How can we help you?</p> 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