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block block-system block-system-breadcrumb-block"> <div class="container"> <nav class="breadcrumb" role="navigation" aria-labelledby="system-breadcrumb"> <ol> <li><a href="https://www.reactionbiology.com">Home</a></li> <li>Services</li><li><a href="https://www.reactionbiology.com/services/target-specific-assays/">Target-specific Assays</a></li><li><a href="https://www.reactionbiology.com/services/kinase-assays/">Kinase Assays</a></li> <li>Kinase SubstrateFinder</li> </ol> </nav> </div> </div> <div class="view-container view-simple-page-conent"> <div class="container"> <article id="post-289" class="post-289 page type-page status-publish hentry" itemtype="https://schema.org/CreativeWork" itemscope=""> <div class="entry-content entry-content-servises "> <h1>Kinase SubstrateFinder Assay Service</h1> <div class="text-mit-bild-text"> <div class="clearfix text-formatted field field--name-field-text field--type-text-long field--label-hidden field__items"> <p>The Kinase SubstrateFinder is a biochemical kinase assay that can be used for the identification of suitable substrates for a specific kinase of interest. Identification of kinase substrates is a necessary step for kinase assay development and is useful for the identification of downstream substrates to explore a pathway.</p> <p>The Kinase SubstrateFinder comprises 5 variants of standardized service packages for the identification of proteins or peptides which can serve as a substrate in biochemical activity assays for a certain protein kinase provided by the customer. The 3 generic Kinase SubstrateFinder variants comprise different panels of proteins used as kinase substrate at Reaction Biology, whereas the physiological SubstrateFinder variants include biotinylated peptide libraries from JPT Peptide Technologies.</p> <ul> <li>Testing of generic substrates with ADP-Glo (Promega) assay format</li> <li>Testing of physiologic substrates with radiometric <sup>33</sup>PanQinase™ assay</li> </ul> </div> <div class="contact-buttons"> <div class="color-scheme-turquoise block block-block-content block-block-content9594f627-1f8b-4f13-b65e-979c19e399c0"> <a href="https://www.reactionbiology.com/contact/contact-us"> <div class="field field--name-field-icon field--type-image field--label-hidden field__items"> <img decoding="async" src="https://www.reactionbiology.com/wp-content/themes/Divi-child/images/icons-email.svg" alt="E-Mail" title="E-Mail" typeof="foaf:Image"> </div> <div class="field field--name-field-text field--type-string field--label-hidden 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alt="Share" title="Share" typeof="foaf:Image"> </div> <div class="field field--name-field-text field--type-string field--label-hidden field__items"> Share </div> </a> </div> </div> </div> </div> </article> </div> </div> <div id="_anchor" class="paragraph weiss black paragraph--type--paragraph-container paragraph--view-mode--default"><div id="ksf-id" class="paragraph paragraph--type--table-one paragraph--view-mode--default"><div class="container center"><h2>List of Substrate Panels</h2> <div class="field field--name-field-table-one field--type-tablefield field--label-hidden field__item"> <div id="tablefield-wrapper-paragraph-2558-field_table_one-0" class="tablefield-wrapper"> <div id="tablefield-paragraph-2558-field_table_one-0_wrapper" class="dataTables_wrapper no-footer"> <table id="tablefield-paragraph-2558-field_table_one-0" class="tablefield dataTable no-footer dtr-inline" role="grid" data-striping="1"> <caption>List of Substrate Panels</caption> <thead> <tr role="row"> <th class="row_0 col_0 all control sorting_asc" tabindex="0" colspan="1" rowspan="1" aria-controls="tablefield-paragraph-2558-field_table_one-0" aria-label="Type of SubstrateFinder: activate to sort column descending" aria-sort="ascending">Type of SubstrateFinder</th> <th class="row_0 col_1 min-fablet sorting" tabindex="0" colspan="1" rowspan="1" aria-controls="tablefield-paragraph-2558-field_table_one-0" aria-label="Protein substrate: activate to sort column ascending">Protein substrate</th> <th class="row_0 col_2 min-fablet sorting" tabindex="0" colspan="1" rowspan="1" aria-controls="tablefield-paragraph-2558-field_table_one-0" aria-label="Applicable for: activate to sort column ascending">Applicable for</th> <th class="row_0 col_3 min-tablet sorting" tabindex="0" colspan="1" rowspan="1" aria-controls="tablefield-paragraph-2558-field_table_one-0" aria-label="Assay system: activate to sort column ascending">Assay system</th> </tr> </thead> <tbody> <tr class="odd" role="row"> <td class="row_2 col_0 all control sorting_1" tabindex="0">Ser/Thr-Generic</td> <td class="row_2 col_1 min-fablet">39</td> <td class="row_2 col_2 min-fablet">Serine/Threonine kinases</td> <td class="row_2 col_3 min-tablet">ADP-Glo assay</td> </tr> <tr class="even" role="row"> <td class="row_3 col_0 all control sorting_1" tabindex="0">Ser/Thr/Tyr-Generic</td> <td class="row_3 col_1 min-fablet">58</td> <td class="row_3 col_2 min-fablet">All protein kinases</td> <td class="row_3 col_3 min-tablet">ADP-Glo assay</td> </tr> <tr class="odd" role="row"> <td class="row_5 col_0 all control sorting_1" tabindex="0">Ser/Thr/Tyr-Physiological</td> <td class="row_5 col_1 min-fablet">720</td> <td class="row_5 col_2 min-fablet">All protein kinases</td> <td class="row_5 col_3 min-tablet">³³PanQinase™</td> </tr> <tr class="even" role="row"> <td class="row_1 col_0 all control sorting_1" tabindex="0">Tyr-Generic</td> <td class="row_1 col_1 min-fablet">19</td> <td class="row_1 col_2 min-fablet">Tyrosine kinases</td> <td class="row_1 col_3 min-tablet">ADP-Glo assay</td> </tr> <tr class="odd" role="row"> <td class="row_4 col_0 all control sorting_1" tabindex="0">Tyr-Physiological</td> <td class="row_4 col_1 min-fablet">145</td> <td class="row_4 col_2 min-fablet">Tyrosine kinases</td> <td class="row_4 col_3 min-tablet">³³PanQinase™</td> </tr> </tbody> </table> </div> </div> </div> </div></div><div id="ksf-id2" class="paragraph paragraph--type--tab-container paragraph--view-mode--default"><div class="container center"><h2>Assay Details</h2> <div class="tab-container"><ul class="tab-navigation"><li class="tab-active" data-id="#tab-ksf-id221">Generic substrate panel</li><li data-id="#tab-ksf-id222">Example, generic substrates</li><li data-id="#tab-ksf-id223">Screening details, generic</li><li data-id="#tab-ksf-id224">Physiological substrate panel</li><li data-id="#tab-ksf-id225">Example, physiological</li><li data-id="#tab-ksf-id226">Screening details, physiological</li></ul><div class="field field--name-field-paragraph field--type-entity-reference-revisions field--label-hidden field__items"><div id="tab-ksf-id221" class="tab-content-active paragraph paragraph--type--tab-text-with-image paragraph--view-mode--default"><div class="field field--name-field-title field--type-string field--label-hidden field__items">Generic substrate panel</div><div class="tab-content"><div class="tab-content-wrapper"><div class="clearfix text-formatted field field--name-field-text field--type-text-long field--label-hidden field__items"><p>Identifying a suitable substrate for a given protein kinase is often a difficult task. Sometimes the physiological substrate of a protein kinase is unknown or does not work under in vitro conditions in a screening assay. Also, some protein kinases will not accept short peptides as substrates but require proteins.</p> <p>Reaction Biology offers a collection of 58 generic serine/threonine- and tyrosine protein kinases substrates that are comprised of proteins, artificial fusion proteins, generic polypeptides, and peptides.</p> <p>Based on our experience our collection of substrates includes at least one that can be identified to be suitable for in vitro kinase assay development.</p> <p>We will verify identified substrates in customized assay setups including the determination of enzymatic parameters V<sub>MAX</sub> and ATP K<sub>m</sub> and/or substrate K<sub>m</sub>.</p> </div></div></div></div><div id="tab-ksf-id222" class=" paragraph paragraph--type--tab-text-with-image paragraph--view-mode--default"><div class="field field--name-field-title field--type-string field--label-hidden field__items">Example, generic substrates</div><div class="tab-content"><div class="tab-content-wrapper"><div class="clearfix text-formatted field field--name-field-text field--type-text-long field--label-hidden field__items"><p><img loading="lazy" decoding="async" class="aligncenter size-full wp-image-1160" src="https://www.reactionbiology.com/wp-content/uploads/2023/07/generic-MELK-example.png" alt="" width="900" height="570" srcset="https://www.reactionbiology.com/wp-content/uploads/2023/07/generic-MELK-example.png 900w, https://www.reactionbiology.com/wp-content/uploads/2023/07/generic-MELK-example-300x190.png 300w, https://www.reactionbiology.com/wp-content/uploads/2023/07/generic-MELK-example-768x486.png 768w" sizes="(max-width: 900px) 100vw, 900px" /></p> <p><strong>Example of MELK assay development with the Generic Kinase SubstrateFinder. </strong></p> <p>MELK kinase activity was measured with a variety of generic substrates. Controls are no-enzyme controls of each substrate and autophosphorylation of the MELK kinase without substrate. Substrate 14 is the most suitable substrate for the establishment of a MELK assay.</p> </div></div></div></div><div id="tab-ksf-id223" class=" paragraph paragraph--type--tab-text-with-image paragraph--view-mode--default"><div class="field field--name-field-title field--type-string field--label-hidden field__items">Screening details, generic</div><div class="tab-content"><div class="tab-content-wrapper"><div class="clearfix text-formatted field field--name-field-text field--type-text-long field--label-hidden field__items"><div class="tab-content"> <div class="tab-content-wrapper"> <div class="clearfix text-formatted field field--name-field-text field--type-text-long field--label-hidden field__items"> <div class="field__item"> <p><strong>Setups: </strong>Singlicate screening with three substrate concentrations.</p> <p><strong>Assay format:</strong> ADP-Glo assay (Promega)</p> <p><strong>ATP concentrations:</strong> 10 µM ATP</p> <p><strong>Controls:</strong> Background and autophosphorylation control. Two control peptides for positive controls provided by the customer.</p> <p><strong>Turnaround time:</strong> 10 business days for standard projects. Expedited scheduling and data delivery can be arranged prior to the commencement of the studies.</p> <p><strong>Report: </strong>The report includes a table of all substrates ranked according to kinase activity, a graphical presentation of the ranking, the identity of the 3 “top scorers”, and a comparison to two control substrates (if provided by the customer).</p> <p><strong>Further testing:</strong> The Kinase SubstrateFinder can be used as a stand-alone service or be part of an assay development package. We can verify identified substrates in customized assay setups including determination of enzymatic parameters V<sub>MAX</sub> and ATP K<sub>m</sub> and/or substrate K<sub>m</sub>.</p> <p><strong>Sample requirements:</strong> In brief, for a standard project, we require 15 µg of the purified kinase. Kinase stock solution concentration should be at least 100 µg/ml.</p> </div> </div> <div class="field field--name-field-pdf field--type-file field--label-hidden field__items"> <div class="field__item"><span class="file file--mime-application-vnd-openxmlformats-officedocument-spreadsheetml-sheet file--x-office-spreadsheet"> <a title="FAQ Assay Condition Form" href="https://www.reactionbiology.com/faq/" target="_blank" rel="noopener">Please see our FAQ page to access our biochemical assay condition form (German facility) for further details on kinase preparation and shipping.</a></span></div> </div> </div> </div> </div></div></div></div><div id="tab-ksf-id224" class=" paragraph paragraph--type--tab-text-with-image paragraph--view-mode--default"><div class="field field--name-field-title field--type-string field--label-hidden field__items">Physiological substrate panel</div><div class="tab-content"><div class="tab-content-wrapper"><div class="clearfix text-formatted field field--name-field-text field--type-text-long field--label-hidden field__items"><p>Your kinase of interest is screened against a library of biotinylated, physiological relevant peptides selected from the SwissProt and Phosphobase databases.<br /> Information you can gain from the Physiological SubstrateFinder service include:</p> <ul> <li>Determination of protein kinase substrate consensus sequences</li> <li>Identification of potential physiological substrates/downstream targets</li> <li>Investigation of signal transduction pathways</li> <li>Identification of protein kinase peptide substrates for screening purposes</li> </ul> </div></div></div></div><div id="tab-ksf-id225" class=" paragraph paragraph--type--tab-text-with-image paragraph--view-mode--default"><div class="field field--name-field-title field--type-string field--label-hidden field__items">Example, physiological</div><div class="tab-content"><div class="tab-content-wrapper"><div class="clearfix text-formatted field field--name-field-text field--type-text-long field--label-hidden field__items"><p><img loading="lazy" decoding="async" class="aligncenter size-full wp-image-1161" src="https://www.reactionbiology.com/wp-content/uploads/2023/07/CDK1-example.png" alt="" width="940" height="417" srcset="https://www.reactionbiology.com/wp-content/uploads/2023/07/CDK1-example.png 940w, https://www.reactionbiology.com/wp-content/uploads/2023/07/CDK1-example-300x133.png 300w, https://www.reactionbiology.com/wp-content/uploads/2023/07/CDK1-example-768x341.png 768w" sizes="(max-width: 940px) 100vw, 940px" /></p> <p><strong>Example of CDK1/CycB1 testing with 720 physiological substrates. </strong></p> <p>Upper left: ranking of all tested peptides from two runs. Upper right: Activity of two control substrates, autophosphorylation (auto) and background (BG) as well as the kinase activity with the 5 best substrates. Table: Listing of the top 5 substrates identified from the substrate library screen.</p> </div></div></div></div><div id="tab-ksf-id226" class=" paragraph paragraph--type--tab-text-with-image paragraph--view-mode--default"><div class="field field--name-field-title field--type-string field--label-hidden field__items">Screening details, physiological</div><div class="tab-content"><div class="tab-content-wrapper"><div class="clearfix text-formatted field field--name-field-text field--type-text-long field--label-hidden field__items"><div class="tab-content"> <div class="tab-content-wrapper"> <div class="clearfix text-formatted field field--name-field-text field--type-text-long field--label-hidden field__items"> <div class="field__item"> <p><strong>Setups:</strong> Single point screening with one peptide concentration.</p> <p><strong>Assay format: </strong>Radiometric <sup>33</sup>PanQinase™ assay</p> <p><strong>ATP concentrations:</strong> 1µM ATP</p> <p><strong>Controls:</strong> Background and autophosphorylation control. Two control peptides for positive controls provided by customer.</p> <p><strong>Turnaround time: </strong>10 business days for standard projects. Expedited scheduling and data delivery can be arranged prior to commencement of the studies.</p> <p><strong>Report:</strong> The raw data and graphical ranking of all peptides is included in the report. Graphical presentation of controls and rank 1 to 5 peptides will be provided beside detailed material and methods.</p> <p><strong>Screening location:</strong> The assay is performed in Freiburg, Germany.</p> <p><strong>Sample requirements:</strong> In brief, for a standard project we require 23 µg of purified kinase. Kinase stock solution concentration should be at least 100 µg/ml.</p> </div> </div> <div class="field field--name-field-pdf field--type-file field--label-hidden field__items"> <div class="field__item"><span class="file file--mime-application-vnd-openxmlformats-officedocument-spreadsheetml-sheet file--x-office-spreadsheet"> <a title="FAQ Assay Condition Form" href="https://www.reactionbiology.com/faq/" target="_blank" rel="noopener">Please see our FAQ page to access our biochemical assay condition form (German facility) for further details on kinase preparation and shipping.</a></span></div> </div> </div> </div> </div></div></div></div></div></div></div></div><div id="ksf-id3" class="paragraph paragraph--type--related-resources-reference paragraph--view-mode--default"><div class="container center"><h2>Related Resources</h2><div class="field--name-field-paragraph related-resources-reference-wrapper"><div class="paragraph--type--related-resource node--type-related-resource"><div 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