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<p><small> <strong>Figure 1. Determination of the antibody titer</strong><br />To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody directed against Flp (cat. No. C15310169) in antigen coated wells. By plotting the absorbance against the antibody dilution (Figure 1), the titer of the antibody was estimated to be 1:9,000.</small></p>
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<p><small> <strong>Figure 2. Western blot analysis using the Diagenode antibody directed against Flp <sup>(1)</sup></strong><br />Western blot was performed on whole cell lysates from untransfected 293 cells (lane 1), or 293 cells transfected with Cre (lane 2), Dre (lane 3) or Flp (lane 4) with the Diagenode antibody against Flp (cat. No. C15310169), diluted 1:500 in BSA/PBS-Tween. The molecular weight marker (in KD) is shown on the left; the location of the protein of interest (expected size: 48 KD) is indicated on the right.</small></p>
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<p>Learn more about: <a href="https://www.diagenode.com/applications/western-blot">Loading control, MW marker visualization</a><em>. <br /></em></p>
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include - APP/View/Products/view.ctp, line 755
View::_evaluate() - CORE/Cake/View/View.php, line 971
View::_render() - CORE/Cake/View/View.php, line 933
View::render() - CORE/Cake/View/View.php, line 473
Controller::render() - CORE/Cake/Controller/Controller.php, line 963
ProductsController::slug() - APP/Controller/ProductsController.php, line 1052
ReflectionMethod::invokeArgs() - [internal], line ??
Controller::invokeAction() - CORE/Cake/Controller/Controller.php, line 491
Dispatcher::_invoke() - CORE/Cake/Routing/Dispatcher.php, line 193
Dispatcher::dispatch() - CORE/Cake/Routing/Dispatcher.php, line 167
[main] - APP/webroot/index.php, line 118
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include - APP/View/Products/view.ctp, line 755
View::_evaluate() - CORE/Cake/View/View.php, line 971
View::_render() - CORE/Cake/View/View.php, line 933
View::render() - CORE/Cake/View/View.php, line 473
Controller::render() - CORE/Cake/Controller/Controller.php, line 963
ProductsController::slug() - APP/Controller/ProductsController.php, line 1052
ReflectionMethod::invokeArgs() - [internal], line ??
Controller::invokeAction() - CORE/Cake/Controller/Controller.php, line 491
Dispatcher::_invoke() - CORE/Cake/Routing/Dispatcher.php, line 193
Dispatcher::dispatch() - CORE/Cake/Routing/Dispatcher.php, line 167
[main] - APP/webroot/index.php, line 118
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<p><small> <strong>Figure 1. Determination of the antibody titer</strong><br />To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody directed against Flp (cat. No. C15310169) in antigen coated wells. By plotting the absorbance against the antibody dilution (Figure 1), the titer of the antibody was estimated to be 1:9,000.</small></p>
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<p><small> <strong>Figure 2. Western blot analysis using the Diagenode antibody directed against Flp <sup>(1)</sup></strong><br />Western blot was performed on whole cell lysates from untransfected 293 cells (lane 1), or 293 cells transfected with Cre (lane 2), Dre (lane 3) or Flp (lane 4) with the Diagenode antibody against Flp (cat. No. C15310169), diluted 1:500 in BSA/PBS-Tween. The molecular weight marker (in KD) is shown on the left; the location of the protein of interest (expected size: 48 KD) is indicated on the right.</small></p>
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include - APP/View/Products/view.ctp, line 755
View::_evaluate() - CORE/Cake/View/View.php, line 971
View::_render() - CORE/Cake/View/View.php, line 933
View::render() - CORE/Cake/View/View.php, line 473
Controller::render() - CORE/Cake/Controller/Controller.php, line 963
ProductsController::slug() - APP/Controller/ProductsController.php, line 1052
ReflectionMethod::invokeArgs() - [internal], line ??
Controller::invokeAction() - CORE/Cake/Controller/Controller.php, line 491
Dispatcher::_invoke() - CORE/Cake/Routing/Dispatcher.php, line 193
Dispatcher::dispatch() - CORE/Cake/Routing/Dispatcher.php, line 167
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<p><small> <strong>Figure 1. Determination of the antibody titer</strong><br />To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody directed against Flp (cat. No. CS-169-100) in antigen coated wells. By plotting the absorbance against the antibody dilution (Figure 1), the titer of the antibody was estimated to be 1:2,000.</small></p>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15310169-WB.png" alt="Western blot" /></p>
</div>
<div class="small-8 columns">
<p><small> <strong>Figure 2. Western blot analysis using the Diagenode antibody directed against Flp</strong><br />Western blot was performed on whole cell lysates from untransfected 293 cells (lane 1), or 293 cells transfected with Cre (lane 2), Dre (lane 3) or Flp (lane 4) with the Diagenode antibody against Flp (cat. No. CS-169-100), diluted 1:500 in BSA/PBS-Tween. The molecular weight marker (in KD) is shown on the left; the location of the protein of interest (expected size: 48 KD) is indicated on the right.</small></p>
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<p>Learn more about: <a href="https://www.diagenode.com/applications/western-blot">Loading control, MW marker visualization</a><em>. <br /></em></p>
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'description' => '<p><strong>Western blot</strong> : The quality of antibodies used in this technique is crucial for correct and specific protein identification. Diagenode offers huge selection of highly sensitive and specific western blot-validated antibodies.</p>
<p>Learn more about: <a href="../applications/western-blot">Loading control, MW marker visualization</a><em>. <br /></em></p>
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'description' => '<p>Cell-to-cell fusion can be quantified by endowing acceptor and donor cells with latent reporter genes/proteins and activators of these genes/proteins, respectively. One way to accomplish this goal is by using a bipartite lentivirus vector (LV)-based cell fusion assay system in which the cellular fusion partners are transduced with a flippase-activatable Photinus pyralis luciferase (PpLuc) expression unit (acceptor cells) or with a recombinant gene encoding FLPeNLS+, a nuclear-targeted and molecularly evolved version of flippase (donor cells). Fusion of both cell populations will lead to the FLPe-dependent generation of a functional PpLuc gene. PpLuc activity is typically measured in cell lysates, precluding consecutive analysis of one cell culture. Therefore, in this study the PpLuc-coding sequence was replaced by that of Gaussia princeps luciferase (GpLuc), a secretory protein allowing repeated analysis of the same cell culture. In myotubes the spread of FLPeNLS+ may be limited due to its nuclear localization signal (NLS) causing low signal outputs. To test this hypothesis, myoblasts were transduced with LVs encoding either FLPeNLS+ or an NLS-less version of FLPe (FLPeNLS-) and subsequently co-cultured in different ratios with myoblasts containing the FLPe-activatable GpLuc expression cassette. At different times after induction of cell-to-cell fusion the GpLuc activity in the culture medium was determined. FLPeNLS+ and FLPeNLS- both activated the latent GpLuc gene but when the percentage of FLPe-expressing myoblasts was limiting, FLPeNLS+ generally yielded slightly higher signals than FLPeNLS- while at low acceptor-to-donor cell ratios FLPeNLS- was usually superior. The ability of FLPeNLS+ to spread through myofibers and to induce reporter gene expression is thus not limited by its NLS. However, at high FLPe concentrations the presence of the NLS negatively affected reporter gene expression. In summary, a rapid and simple chemiluminescence assay for quantifying cell-to-cell fusion progression based on GpLuc has been developed.</p>',
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View::_evaluate() - CORE/Cake/View/View.php, line 971
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×