Isotype Control

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mb-4">Isotype Control</h1> <p class="large"> Antibodies bind to a specific protein or antigen, but sometimes an antibody can bind to other proteins or cellular components in a non-specific manner. Experimental controls are critical in determining if the antibodies used in your experiment have any off-target binding capabilities. <br /><br />Off-target or non-specific binding by antibodies lead to false positive results, making it difficult to determine background staining from antigen-specific staining. Therefore, it is important to always include experimental controls to ensure reliable results. </p> <a href="/results?q=isotype+control" class="btn btn-primary mt-4 mt-lg-3 mb-4">View all Isotype Controls</a> </div> </div> </div> </div> </section> <section class="landing__content bg-ptg-grey-6"> <div class="container-xl"> <div class="row"> <div class="col-12 col-lg-2 mb-3"> <div class="in-page-nav" style="top: 170px"> <h5>Jump to:</h5> <ul class="list-unstyled"> <li><a href="#what">What is an isotype control?</a></li> <li><a href="#why">Why use an isotype control?</a></li> <li><a href="#how">How do I choose the best isotype control?</a></li> <li><a href="#which">Which applications use isotype controls?</a></li> <li><a href="#products">Proteintech Isotype Control Products</a></li> </ul> </div> </div> <div class="col-12 col-lg-9"> <div class="row"> <div class="col-12 col-lg-12"> <div id="what"></div> <h3 class="ptg-darkblue mb-4">What is an isotype control? </h3> <p class="mb-5"> An isotype control is an antibody that is used as a negative control in antibody staining applications. <br /><br />Isotype control antibodies have the same immunoglobulin class and subclass as the primary antibody being used in the experiment but lack specificity to the protein of interest. </p> <img class="mb-5" width="100%" src="https://www.ptglab.com/media/6174/isotype_control_conjugated_primary_antibody.png" alt="isotype control conjugated primary antibody" /> </div> </div> <div class="row"> <div class="col-12"> <div id="why"></div> <h3 class="ptg-darkblue mb-4"> Why use an isotype control? </h3> </div> </div> <p class="mb-5"> Isotype controls allow you to distinguish between specific antibody staining and background staining in your experiments. Antibodies (and sometimes their conjugates) can non-specifically bind to other proteins, endogenous enzymes, lipids, and Fc receptors. Including an isotype control as a negative control in your experiment also allows you to determine whether the blocking and/or washing steps in your protocol are sufficient. </p> <div class="row"> <div class="col-12"> <div id="how"></div> <h3 class="ptg-darkblue mb-4"> How do I choose the best isotype control? </h3> </div> </div> <p class="mb-3"> An isotype control should match certain characteristics of your primary antibody: </p> <div class="row"> <div class="col-12 col-lg-4"> <ul class="list-unstyled"> <li> <div class="d-flex align-items-center gap-3 mb-3"> <div class="box box-sm bg-ptg-lightblue"> <img src="/img/icons/Icon_Tick-white.svg" width="50"> </div> <p>Same host species<p> </div> </li> <li> <div class="d-flex align-items-center gap-3 mb-3"> <div class="box box-sm bg-ptg-lightblue"> <img src="/img/icons/Icon_Tick-white.svg" width="50"> </div> <p>Same Ig class </p> </div> </li> </ul> </div> <div class="col-12 col-lg-6 mb-4"> <div class="col-md-9"> <ul class="list-unstyled"> <li> <div class="d-flex align-items-center gap-3 mb-3"> <div class="box box-sm bg-ptg-lightblue"> <img src="/img/icons/Icon_Tick-white.svg" width="50"> </div> <p>Same Ig subclass (for some species) </p> </div> </li> <li> <div class="d-flex align-items-center gap-3 mb-3"> <div class="box box-sm bg-ptg-lightblue"> <img src="/img/icons/Icon_Tick-white.svg" width="50"> </div> <p>Same label as primary antibody (biotin, HRP, fluorophore, etc.)<p> </div> </li> </ul> </div> </div> </div> <p> As an example, if you are using a primary antibody that is a mouse IgG2b antibody conjugated to FITC, you would want to choose a mouse IgG2b isotype control antibody that is also conjugated to FITC. </p> <p class="mb-5"> If you are using an unconjugated primary antibody and a secondary antibody for detection, your isotype control should match your primary antibody to determine non-specific binding of the primary antibody. A secondary-only control (a sample stained with only secondary antibody; no primary antibody used) can be included as a control to determine any background staining by the secondary antibody. </p> <img class="mb-5" width="100%" src="https://www.ptglab.com/media/6175/isotype_control_unconjugated_primary_antibody_with_secondary_antibody.png?" alt="isotype control unconjugated primary antibody with secondary antibody" /> <div class="row"> <div class="col-12 col-lg-12"> <div id="which"></div> <h3 class="ptg-darkblue mb-4">Which applications use isotype controls?</h3> <p class="mb-4"> Isotype controls are most commonly used in flow cytometry, immunoprecipitation and IHC, but can also be used for blocking and coating purposes in other applications such as WB, ELISA, and IF/ICC. </p> <h5>Protocol for isotype controls</h5> <p> The isotype control antibody simply replaces the primary antibody for the control samples, and the rest of the protocol is exactly the same (this eliminates any other experimental variables). </p> <p> The isotype controls should be handled under the exact same conditions as the experimental samples; this includes using the isotype control antibody at the same concentration as the primary antibody. </p> <p> If a secondary antibody is used, the isotype controls should be stained with the same secondary antibody. </p> <p class="mb-4"> When using isotype controls your data should look similar to the example below. Using rabbit IgG isotype control recombinant antibody (98136-1-RR) makes it easier to gate the positive population of CD4+ cells that are specifically stained with the anti-human CD4 rabbit recombinant antibody (98042-1-RR). </p> <div class="col-12 mb-4"> <a href="/products/IgG-Isotype-Control-Antibody-98136-1-RR.htm" target="_blank"><img width="100%" src="/media/wwwnzb4f/isotype20241015.jpg" alt="Flow cytometry validation image. 1X10^6 human peripheral blood lymphocytes were surface stained with 0.06 ug APC Mouse IgG2b Isotype Control (APC-65128, clone: MPC-11) (red) or 0.06 ug APC Anti-Human CD4 (APC-65134, clone: OKT4) (orange), or not stained (blank) (blue). Cells were not fixed."></a> </div> <div class="col-12 mb-4"> <span>1x10^6 human PBMCs were surface stained with 0.2 ug Rabbit IgG Isotype Control Recombinant Antibody (98136-1-RR, Clone: 240953C9) and PE-Conjugated Goat Anti-Rabbit IgG(H+L) (red in histogram), or 0.2 ug Anti-Human CD4 Rabbit Recombinant Antibody (98042-1-RR) and PE-Conjugated Goat Anti-Rabbit IgG(H+L) (green in histogram), or unstained (blue in histogram). Cells were not fixed.</span> </div> <div id="products"></div> <h5>Proteintech Isotype Control Products</h5> <ul class="mb-4"> <li>More than 100 isotype control antibodies</li> <li>Available as unconjugated, or conjugated to: APC, FITC, PE, CoraLite®Plus 405, CoraLite®Plus 488, CoraLite®Plus 555, CoraLite®594, CoraLite® Plus 647, CoraLite® Plus 750, Atlantic Blue™, Cardinal Red™</li> <li>Available species: Mouse, Rat, Rabbit, Armenian Hamster</li> </ul> <h5>Comparison Table of Proteintech Isotype Controls by Sub-class</h5> </div> </div> <div> <div class="row"> <div class="col-lg-12"> <table class="table"> <thead> <tr> <th scope="col"><strong>Species</strong></th> <th scope="col"><strong>Subclass</strong></th> <th scope="col"><strong>Isotype Control</strong></th> </tr> </thead> <tbody> <tr> <td>Rabbit</td> <td>IgG</td> <td><a href="/results?q=Rabbit%20IgG%20isotype%20control&filter=Clonality%3DRecombinant&mode=">Rabbit IgG Isotype Control Recombinant Antibody</a></td> </tr> <tr> <td>Rabbit</td> <td>IgG</td> <td><a href="/products/IgG-control-Antibody-30000-0-AP.htm">Rabbit IgG Isotype Control Polyclonal Antibody</a></td> </tr> <tr> <td>Mouse</td> <td>IgG1</td> <td><a href="/results?q=IgG1%20Isotype%20Control&filter=Host%20Species%3DMouse&mode=">Mouse IgG1 Isotype Control</a></td> </tr> <tr> <td>Mouse</td> <td>IgG2a</td> <td><a href="/results?q=IgG2a%20Isotype%20Control&filter=Host%20Species%3DMouse&mode=">Mouse IgG2a Isotype Control</a></td> </tr> <tr> <td>Mouse</td> <td>IgG2b</td> <td><a href="/results?q=IgG2b%20Isotype%20Control&filter=Host%20Species%3DMouse&mode=">Mouse IgG2b Isotype Control</a></td> </tr> <tr> <td>Rat</td> <td>IgG1</td> <td><a href="/results?q=IgG1%20Isotype%20Control&filter=Host%20Species%3DRat&mode=">Rat IgG1 Isotype Control</a></td> </tr> <tr> <td>Rat</td> <td>IgG2a</td> <td><a href="/results?q=IgG2a%20Isotype%20Control&filter=Host%20Species%3DRat&mode=">Rat IgG2a Isotype Control</a></td> </tr> <tr> <td>Rat</td> <td>IgG2b</td> <td><a href="/results?q=IgG2b%20Isotype%20Control&filter=Host%20Species%3DRat&mode=">Rat IgG2b Isotype Control</a></td> </tr> <tr> <td>Hamster</td> <td>IgG</td> <td><a href="/results?q=IgG%20Isotype%20Control&filter=Host%20Species%3DArmenian%20Hamster&mode=">Armenian Hamster IgG Isotype Control</a></td> </tr> </tbody> </table> </div> </div> <div class="row justify-content-center"> <div class="col-12 col-lg-auto"> <a href="/results?q=isotype+control" class="btn btn-md text-nowrap d-none d-lg-inline-block btn-primary">View all Isotype Controls</a> <a href="/results?q=isotype+control" class="btn btn-md text-nowrap d-inline-block d-lg-none w-100 btn-primary mb-3">View all Isotype Controls</a> </div> </div> </div> </div> </div> </div> </section> <div class="scroll-to-top"> <a href="#top" title="Back to Top" class="btn btn-roundel"> <svg xmlns="http://www.w3.org/2000/svg" fill="currentColor" viewBox="0 0 16 16"> <path fill-rule="evenodd" d="M8 15a.5.5 0 0 0 .5-.5V2.707l3.146 3.147a.5.5 0 0 0 .708-.708l-4-4a.5.5 0 0 0-.708 0l-4 4a.5.5 0 1 0 .708.708L7.5 2.707V14.5a.5.5 0 0 0 .5.5z"></path> </svg> </a> </div> <footer class="footer"> <div class="container-xl"> <div class="row d-flex d-lg-none"> <div class="col-12 text-center"> <hr> <ul class="inline-list m-0 p-0 opacity-7"> <li class="list-inline-item"> <a href="http://www.youtube.com/proteintech"> <img loading="lazy" src="/img/logos/Icon_Youtube.svg" width="42"> </a> </li> <li class="list-inline-item"> <a href="https://www.linkedin.com/company/proteintech-group/"> <img loading="lazy" src="/img/logos/Icon_Linkedin.svg" width="42"> </a> </li> <li class="list-inline-item"> <a href="http://www.twitter.com/proteintech"> <img loading="lazy" src="/img/logos/Icon_Twitter.svg" width="42"> </a> </li> <li class="list-inline-item"> <a href="https://www.facebook.com/Proteintech-311515338877884/timeline/"> <img loading="lazy" src="/img/logos/Icon_Facebook.svg" width="42"> </a> </li> <li class="list-inline-item"> <a href="https://www.instagram.com/proteintech/"> <img loading="lazy" src="/img/logos/Icon_Instagram.svg" width="42"> </a> </li> <li class="list-inline-item ms-2"> <a name="livechat"> <img loading="lazy" src="/img/icons/Icon_Chat_Black.png" width="30"> </a> </li> </ul> <hr> <p class="fw-semi m-4"> <a class="mx-2" href="/contact/">Contact Us</a> <a class="mx-2" href="/news/blog/privacy-policy/">Privacy Policy</a> <a class="mx-2" href="/terms-of-use/">Terms of use</a> <a class="mx-2" href="/news/blog/product-terms/">Terms and Conditions</a> <a class="mx-2" href="/trademark-information/">Trademark Information</a> <a class="mx-2" href="/imprint-impressum/" target="_blank">Imprint (Impressum)</a> <a class="mx-2" href="/modern-slavery-statement/">Modern Slavery Statement</a> <a class="mx-2" href="/chromotek-privacy-policy/">ChromoTek GmbH Privacy Policy</a> </p> <p class="fw-semi"> Copyright © 2002-2022 Proteintech Group, Inc. 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