{"id":2930,"date":"2024-07-01T20:11:26","date_gmt":"2024-07-01T20:11:26","guid":{"rendered":"https:\/\/praxilabs.com\/en\/blog\/?p=2930"},"modified":"2025-08-22T20:50:39","modified_gmt":"2025-08-22T20:50:39","slug":"polyacrylamide-gel-electrophoresis","status":"publish","type":"post","link":"https:\/\/praxilabs.com\/en\/blog\/2024\/07\/01\/polyacrylamide-gel-electrophoresis\/","title":{"rendered":"Exploring Polyacrylamide Gel Electrophoresis (PAGE) | Your Essential Q&#038;A Guide"},"content":{"rendered":"<p><span style=\"font-weight: 400; font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\">Polyacrylamide Gel Electrophoresis (PAGE) is a fundamental technique in the realm of molecular biology, enabling the separation and analysis of macromolecules such as DNA, RNA, and proteins based on their size and charge. As a cornerstone of scientific research and clinical diagnostics, understanding the intricacies of this method is essential.<\/span><\/p>\n<p><span style=\"font-weight: 400; font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\">\u00a0In this blog, we&#8217;ll delve into the world of Polyacrylamide Gel Electrophoresis, exploring common questions and providing insightful answers to demystify this powerful tool.<\/span><\/p>\n<div id=\"ez-toc-container\" class=\"ez-toc-v2_0_82_2 counter-hierarchy ez-toc-counter ez-toc-light-blue ez-toc-container-direction\">\r\n<div class=\"ez-toc-title-container\">\r\n<p class=\"ez-toc-title\" style=\"cursor:inherit\">Table of Contents<\/p>\r\n<span class=\"ez-toc-title-toggle\"><\/span><\/div>\r\n<nav><ul class='ez-toc-list ez-toc-list-level-1 ' ><li class='ez-toc-page-1 ez-toc-heading-level-2'><a class=\"ez-toc-link ez-toc-heading-1\" href=\"https:\/\/praxilabs.com\/en\/blog\/2024\/07\/01\/polyacrylamide-gel-electrophoresis\/#What_is_the_Polyacrylamide_Gel_Electrophoresis_PAGE\" >What is the Polyacrylamide Gel Electrophoresis PAGE?<\/a><\/li><li class='ez-toc-page-1 ez-toc-heading-level-2'><a class=\"ez-toc-link ez-toc-heading-2\" href=\"https:\/\/praxilabs.com\/en\/blog\/2024\/07\/01\/polyacrylamide-gel-electrophoresis\/#What_is_the_SDS-PAGE_of_a_protein\" >What is the SDS-PAGE of a protein?<\/a><\/li><li class='ez-toc-page-1 ez-toc-heading-level-2'><a class=\"ez-toc-link ez-toc-heading-3\" href=\"https:\/\/praxilabs.com\/en\/blog\/2024\/07\/01\/polyacrylamide-gel-electrophoresis\/#How_does_polyacrylamide_gel_electrophoresis_PAGE_work\" >How does polyacrylamide gel electrophoresis (PAGE) work?<\/a><\/li><li class='ez-toc-page-1 ez-toc-heading-level-2'><a class=\"ez-toc-link ez-toc-heading-4\" href=\"https:\/\/praxilabs.com\/en\/blog\/2024\/07\/01\/polyacrylamide-gel-electrophoresis\/#What_does_the_SDS-PAGE_tell_you\" >What does the SDS-PAGE tell you?<\/a><\/li><li class='ez-toc-page-1 ez-toc-heading-level-2'><a class=\"ez-toc-link ez-toc-heading-5\" href=\"https:\/\/praxilabs.com\/en\/blog\/2024\/07\/01\/polyacrylamide-gel-electrophoresis\/#Why_do_we_use_PAGE_for_protein_electrophoresis\" >Why do we use PAGE for protein electrophoresis?<\/a><ul class='ez-toc-list-level-3' ><li class='ez-toc-heading-level-3'><a class=\"ez-toc-link ez-toc-heading-6\" href=\"https:\/\/praxilabs.com\/en\/blog\/2024\/07\/01\/polyacrylamide-gel-electrophoresis\/#Applications_of_Protein_Gel_Electrophoresis\" >Applications of Protein Gel Electrophoresis:<\/a><\/li><\/ul><\/li><li class='ez-toc-page-1 ez-toc-heading-level-2'><a class=\"ez-toc-link ez-toc-heading-7\" href=\"https:\/\/praxilabs.com\/en\/blog\/2024\/07\/01\/polyacrylamide-gel-electrophoresis\/#Discover_Polyacrylamide_Gel_Electrophoresis_Virtual_labs_from_PraxiLabs\" >Discover Polyacrylamide Gel Electrophoresis Virtual labs from PraxiLabs\u00a0<\/a><ul class='ez-toc-list-level-3' ><li class='ez-toc-heading-level-3'><a class=\"ez-toc-link ez-toc-heading-8\" href=\"https:\/\/praxilabs.com\/en\/blog\/2024\/07\/01\/polyacrylamide-gel-electrophoresis\/#_Steps_of_Polyacrylamide_Gel_Electrophoresis_Simulation\" >\u00a0Steps of Polyacrylamide Gel Electrophoresis Simulation<\/a><ul class='ez-toc-list-level-4' ><li class='ez-toc-heading-level-4'><a class=\"ez-toc-link ez-toc-heading-9\" href=\"https:\/\/praxilabs.com\/en\/blog\/2024\/07\/01\/polyacrylamide-gel-electrophoresis\/#First_Gel_and_buffer_preparation\" >First: Gel and buffer preparation<\/a><\/li><li class='ez-toc-page-1 ez-toc-heading-level-4'><a class=\"ez-toc-link ez-toc-heading-10\" href=\"https:\/\/praxilabs.com\/en\/blog\/2024\/07\/01\/polyacrylamide-gel-electrophoresis\/#Second_steps_of_gel_casting_and_sample_application\" >Second: steps of gel casting and sample application<\/a><\/li><\/ul><\/li><li class='ez-toc-page-1 ez-toc-heading-level-3'><a class=\"ez-toc-link ez-toc-heading-11\" href=\"https:\/\/praxilabs.com\/en\/blog\/2024\/07\/01\/polyacrylamide-gel-electrophoresis\/#General_Guidelines_for_SDS-PAGE_SDS-_Polyacrylamide_Gel_Electrophoresis_Experiment\" >General Guidelines for SDS-PAGE (SDS- Polyacrylamide Gel Electrophoresis) Experiment<\/a><\/li><\/ul><\/li><\/ul><\/nav><\/div>\r\n<h2><span class=\"ez-toc-section\" id=\"What_is_the_Polyacrylamide_Gel_Electrophoresis_PAGE\"><\/span><span style=\"font-size: 18pt; font-family: tahoma, arial, helvetica, sans-serif;\"><b>What is the Polyacrylamide Gel Electrophoresis PAGE?<\/b><\/span><span class=\"ez-toc-section-end\"><\/span><\/h2>\n<p><span style=\"font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\">Polyacrylamide Gel Electrophoresis (PAGE) is an <a href=\"https:\/\/www.sciencedirect.com\/science\/article\/abs\/pii\/B9780444595621000104\" target=\"_blank\" rel=\"noopener\">electrophoretic technique<\/a> that is used mainly for protein analysis by separating proteins, nucleic acid fragments (smaller than 100bp), and polypeptide chains depending on their electrophoretic mobility. It is widely used in molecular biology, biochemistry, genetics, forensic chemistry, and biotechnology.<\/span><\/p>\n<p><span style=\"font-family: tahoma, arial, helvetica, sans-serif;\"><img loading=\"lazy\" decoding=\"async\" class=\"alignnone size-full wp-image-2936\" src=\"https:\/\/praxilabs.com\/en\/blog\/wp-content\/uploads\/2024\/07\/SDS-Polyacrylamide-Gel-Electrophoresis.jpg\" alt=\"What is the Polyacrylamide Gel Electrophoresis PAGE?\" width=\"1200\" height=\"628\" srcset=\"https:\/\/praxilabs.com\/en\/blog\/wp-content\/uploads\/2024\/07\/SDS-Polyacrylamide-Gel-Electrophoresis.jpg 1200w, https:\/\/praxilabs.com\/en\/blog\/wp-content\/uploads\/2024\/07\/SDS-Polyacrylamide-Gel-Electrophoresis-300x157.jpg 300w, https:\/\/praxilabs.com\/en\/blog\/wp-content\/uploads\/2024\/07\/SDS-Polyacrylamide-Gel-Electrophoresis-1024x536.jpg 1024w, https:\/\/praxilabs.com\/en\/blog\/wp-content\/uploads\/2024\/07\/SDS-Polyacrylamide-Gel-Electrophoresis-768x402.jpg 768w\" sizes=\"auto, (max-width: 1200px) 100vw, 1200px\" \/><\/span><\/p>\n<h2><span class=\"ez-toc-section\" id=\"What_is_the_SDS-PAGE_of_a_protein\"><\/span><span style=\"font-size: 18pt; font-family: tahoma, arial, helvetica, sans-serif;\"><b>What is the SDS-PAGE of a protein?<\/b><\/span><span class=\"ez-toc-section-end\"><\/span><\/h2>\n<p><span style=\"font-weight: 400; font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\">SDS-PAGE (SDS-polyacrylamide gel electrophoresis) is an electrophoretic technique that separates polypeptide chains according to their molecular weights (Mr). This method uses polyacrylamide gel containing sodium dodecyl sulfate (SDS) which is added to the system in order to provide denaturing conditions.<\/span><\/p>\n<p><span style=\"font-weight: 400; font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\">You can tell from its name that the technique utilizes polyacrylamide gel containing sodium dodecyl sulfate (SDS). SDS in the sample and the gel cancels the effect of intrinsic electrical charge of the sample proteins. All proteins acquire a negatively charged rod like structure, so separation becomes largely dependent on Molecular weight of sample proteins.<\/span><\/p>\n<p><span style=\"font-weight: 400; font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\">\u00a0The gel used in SDS PAGE can be divided into stacking gel and separating gel. Stacking gel (acrylamide 5%) is poured on top of the separating gel (after solidification) and a gel comb is inserted in the stacking gel. The acrylamide percentage is chosen in accordance with the size of target proteins in the sample.<\/span><\/p>\n<p><span style=\"font-family: tahoma, arial, helvetica, sans-serif;\"><img loading=\"lazy\" decoding=\"async\" class=\"alignnone size-full wp-image-2937\" src=\"https:\/\/praxilabs.com\/en\/blog\/wp-content\/uploads\/2024\/07\/Polyacrylamide-Gel-Electrophoresis-PAGE.jpg\" alt=\"How does polyacrylamide gel electrophoresis (PAGE) work?\" width=\"1200\" height=\"628\" srcset=\"https:\/\/praxilabs.com\/en\/blog\/wp-content\/uploads\/2024\/07\/Polyacrylamide-Gel-Electrophoresis-PAGE.jpg 1200w, https:\/\/praxilabs.com\/en\/blog\/wp-content\/uploads\/2024\/07\/Polyacrylamide-Gel-Electrophoresis-PAGE-300x157.jpg 300w, https:\/\/praxilabs.com\/en\/blog\/wp-content\/uploads\/2024\/07\/Polyacrylamide-Gel-Electrophoresis-PAGE-1024x536.jpg 1024w, https:\/\/praxilabs.com\/en\/blog\/wp-content\/uploads\/2024\/07\/Polyacrylamide-Gel-Electrophoresis-PAGE-768x402.jpg 768w\" sizes=\"auto, (max-width: 1200px) 100vw, 1200px\" \/><\/span><\/p>\n<h2><span class=\"ez-toc-section\" id=\"How_does_polyacrylamide_gel_electrophoresis_PAGE_work\"><\/span><span style=\"font-size: 18pt; font-family: tahoma, arial, helvetica, sans-serif;\"><b>How does polyacrylamide gel electrophoresis (PAGE) work?<\/b><\/span><span class=\"ez-toc-section-end\"><\/span><\/h2>\n<p><span style=\"font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\">The main principle of Polyacrylamide Gel Electrophoresis PAGE is to separate molecules (proteins, or polypeptides, or nucleic acids) by passing them through the pores of a polyacrylamide gel using an electric current. To achieve this, an acrylamide\u2013bisacrylamide mix is polymerized (polyacrylamide) by the addition of ammonium persulfate (APS).<\/span><\/p>\n<p><span style=\"font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\">The reaction, which is catalyzed by tetramethylethylenediamine (TEMED), forms a net-like structure with pores through which analytes can move. The higher the percentage of total acrylamide included in the gel, the smaller the pore size, hence the smaller the proteins that will be able to pass through. The ratio of acrylamide to bisacrylamide will also impact pore size but this is often kept constant. Smaller pore sizes also reduce the speed at which small proteins are able to move through the gel, improving their resolution and preventing them from running off into the buffer rapidly when current is applied.<\/span><\/p>\n<p><span style=\"font-family: tahoma, arial, helvetica, sans-serif;\"><a href=\"https:\/\/www.technologynetworks.com\/analysis\/articles\/polyacrylamide-gel-electrophoresis-how-it-works-technique-variants-and-its-applications-359100#:~:text=1)%20Samples%20are%20prepared%20for,gels%20are%20stained%20and%20visualized\" target=\"_blank\" rel=\"noopener\"><span style=\"font-size: 10pt;\"><strong>Source<\/strong><\/span><\/a><\/span><\/p>\n<p><span style=\"font-family: tahoma, arial, helvetica, sans-serif;\"><span style=\"font-size: 10pt;\"><i>Polyacrylamide gel electrophoresis, how it works, technique variants and its applications<\/i>, <i>Analysis &amp; Separations from Technology Networks<\/i>. Available at:\u00a0 (Accessed:01 July 2024).<\/span><\/span><\/p>\n<h2><span class=\"ez-toc-section\" id=\"What_does_the_SDS-PAGE_tell_you\"><\/span><span style=\"font-size: 18pt; font-family: tahoma, arial, helvetica, sans-serif;\"><b>What does the SDS-PAGE tell you?<\/b><\/span><span class=\"ez-toc-section-end\"><\/span><\/h2>\n<p><span style=\"font-weight: 400; font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\">The SDS-PAGE provides us with information on the mass, charge, purity, or presence of a protein by separating polypeptide chains according to their molecular weights.<\/span><\/p>\n<p><span style=\"font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\"><b>In PraxiLabs, we provide several <a href=\"https:\/\/praxilabs.com\">virtual labs simulations<\/a> for protein electrophoresis such as:<\/b><\/span><\/p>\n<ul>\n<li><span style=\"font-family: tahoma, arial, helvetica, sans-serif;\"><strong><a href=\"https:\/\/praxilabs.com\/en\/3d-simulations\/protein-electrophoresis-virtual-lab-simulation\"><span style=\"font-size: 14pt;\">Protein Electrophoresis<\/span><\/a><span style=\"font-size: 14pt;\"> (Polyacrylamide Gel Electrophoresis &#8211; PAGE)<\/span><\/strong><\/span><\/li>\n<\/ul>\n<p><span style=\"font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\">Learn polyacrylamide gel electrophoresis protocol and practice proper sample preparation for SDS-PAGE (SDS-polyacrylamide gel electrophoresis.<\/span><\/p>\n<ul>\n<li><span style=\"font-family: tahoma, arial, helvetica, sans-serif;\"><strong><a href=\"https:\/\/praxilabs.com\/EnglishContent\/Final_Experiments_PDF\/Biology\/Molecular-Biology\/2d-protein-electrophoresis-simulation.pdf\"><span style=\"font-size: 14pt;\">2D Protein Electrophoresis<\/span><\/a><span style=\"font-size: 14pt;\"> (Isoelectric Point Focusing, PAGE)<\/span><\/strong><\/span><\/li>\n<\/ul>\n<p><span style=\"font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\">Learn how to extract the cellular proteome from a sample, followed by its isoelectric focusing and SDS-polyacrylamide gel electrophoresis.<\/span><\/p>\n<ul>\n<li><span style=\"font-family: tahoma, arial, helvetica, sans-serif;\"><strong><a href=\"https:\/\/praxilabs.com\/en\/3d-simulations\/western-blot-biology-virtual-lab-simulation\"><span style=\"font-size: 14pt;\">Western Blot<\/span><\/a><\/strong><\/span><\/li>\n<\/ul>\n<p><span style=\"font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\">Understand how to detect a specific protein in a sample using electroblotting of proteins into nitrocellulose membrane.<\/span><\/p>\n<ul>\n<li><span style=\"font-family: tahoma, arial, helvetica, sans-serif;\"><strong><a href=\"https:\/\/praxilabs.com\/en\/3d-simulations\/agarose-gel-electrophoresis-virtual-lab-simulation\"><span style=\"font-size: 14pt;\">Agarose Gel Electrophoresis<\/span><\/a><\/strong><\/span><\/li>\n<\/ul>\n<p><span style=\"font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\">Learn how to separate and identify DNA or RNA molecules by size, using an electric current.<\/span><\/p>\n<p style=\"text-align: center;\"><iframe loading=\"lazy\" src=\"\/\/www.youtube.com\/embed\/k9Kmvh9blog\" width=\"560\" height=\"314\" allowfullscreen=\"allowfullscreen\"><\/iframe><\/p>\n<p style=\"text-align: center;\"><span style=\"font-size: 18pt; font-family: tahoma, arial, helvetica, sans-serif;\"><b>\u00a0<\/b><a href=\"https:\/\/praxilabs.com\/en\/pricing\"><b>Subscribe and Try them Now!<\/b><\/a><\/span><\/p>\n<h2><span class=\"ez-toc-section\" id=\"Why_do_we_use_PAGE_for_protein_electrophoresis\"><\/span><span style=\"font-size: 18pt; font-family: tahoma, arial, helvetica, sans-serif;\"><b>Why do we use PAGE for protein electrophoresis?<\/b><\/span><span class=\"ez-toc-section-end\"><\/span><\/h2>\n<p><span style=\"font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\"><b>We use PAGE for protein electrophoresis<\/b><span style=\"font-weight: 400;\"> to obtain high resolution separation of complex mixtures of proteins by separating polypeptide chains according to their molecular weights.<\/span><\/span><\/p>\n<h3><span class=\"ez-toc-section\" id=\"Applications_of_Protein_Gel_Electrophoresis\"><\/span><span style=\"font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\"><b>Applications of Protein Gel Electrophoresis:<\/b><\/span><span class=\"ez-toc-section-end\"><\/span><\/h3>\n<p><span style=\"font-weight: 400; font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\">Once proteins have been separated by gel electrophoresis, they can be utilized for a number of downstream applications including:<\/span><\/p>\n<ul>\n<li><span style=\"font-weight: 400; font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\"> Determining the size and IP of proteins.<\/span><\/li>\n<li><span style=\"font-weight: 400; font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\"> Enzymatic assays.<\/span><\/li>\n<li><span style=\"font-weight: 400; font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\"> Further purification.<\/span><\/li>\n<li><span style=\"font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\"> Transfer to a membrane for immunological detection (immunoblotting or <a href=\"https:\/\/praxilabs.com\/en\/3d-simulations\/western-blot-biology-virtual-lab-simulation\">western blot<\/a>).<\/span><\/li>\n<li><span style=\"font-weight: 400; font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\"> Elution and digestion for mass spectrometric analysis.<\/span><\/li>\n<li><span style=\"font-weight: 400; font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\"> Serum protein electrophoresis is conducted when a patient has an abnormal result on a total protein or albumin blood test or has symptoms of diseases that are associated with abnormal protein production, such as multiple myeloma or multiple sclerosis.<\/span><\/li>\n<\/ul>\n<p style=\"text-align: center;\"><span style=\"font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\"><b>Ready to revolutionize your biology education? Dive into PraxiLabs&#8217; virtual labs and gain unlimited access to cutting-edge experiments<\/b><\/span><\/p>\n<p style=\"text-align: center;\"><span style=\"font-family: tahoma, arial, helvetica, sans-serif;\"><a href=\"https:\/\/praxilabs.com\/en\/sign-up\"><b>Join Praxilabs For Free Now!<\/b><\/a><\/span><\/p>\n<p><span style=\"font-family: tahoma, arial, helvetica, sans-serif;\"><img loading=\"lazy\" decoding=\"async\" class=\"alignnone size-full wp-image-2933\" src=\"https:\/\/praxilabs.com\/en\/blog\/wp-content\/uploads\/2024\/07\/Discover-Polyacrylamide-Gel-Electrophoresis-Virtual-labs-from-PraxiLabs.jpg\" alt=\"Discover Polyacrylamide Gel Electrophoresis Virtual labs from PraxiLabs\u00a0\" width=\"1200\" height=\"628\" srcset=\"https:\/\/praxilabs.com\/en\/blog\/wp-content\/uploads\/2024\/07\/Discover-Polyacrylamide-Gel-Electrophoresis-Virtual-labs-from-PraxiLabs.jpg 1200w, https:\/\/praxilabs.com\/en\/blog\/wp-content\/uploads\/2024\/07\/Discover-Polyacrylamide-Gel-Electrophoresis-Virtual-labs-from-PraxiLabs-300x157.jpg 300w, https:\/\/praxilabs.com\/en\/blog\/wp-content\/uploads\/2024\/07\/Discover-Polyacrylamide-Gel-Electrophoresis-Virtual-labs-from-PraxiLabs-1024x536.jpg 1024w, https:\/\/praxilabs.com\/en\/blog\/wp-content\/uploads\/2024\/07\/Discover-Polyacrylamide-Gel-Electrophoresis-Virtual-labs-from-PraxiLabs-768x402.jpg 768w\" sizes=\"auto, (max-width: 1200px) 100vw, 1200px\" \/><\/span><\/p>\n<h2><span class=\"ez-toc-section\" id=\"Discover_Polyacrylamide_Gel_Electrophoresis_Virtual_labs_from_PraxiLabs\"><\/span><span style=\"font-size: 18pt; font-family: tahoma, arial, helvetica, sans-serif;\"><b>Discover Polyacrylamide Gel Electrophoresis Virtual labs from PraxiLabs\u00a0<\/b><\/span><span class=\"ez-toc-section-end\"><\/span><\/h2>\n<p><span style=\"font-weight: 400; font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\">By the end of this simulation students should be able to:<\/span><\/p>\n<ul>\n<li style=\"font-weight: 400;\" aria-level=\"1\"><span style=\"font-weight: 400; font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\">\u00a0Prepare polyacrylamide gel properly according to the size of target proteins. Interpret the results of a successful protein electrophoresis run.<\/span><\/li>\n<li style=\"font-weight: 400;\" aria-level=\"1\"><span style=\"font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\"><span style=\"font-weight: 400;\">\u00a0<\/span><span style=\"font-weight: 400;\">Learn polyacrylamide gel electrophoresis protocol.<\/span><\/span><\/li>\n<li style=\"font-weight: 400;\" aria-level=\"1\"><span style=\"font-weight: 400; font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\">Practice proper sample preparation for SDS-PAGE (SDS-polyacrylamide gel electrophoresis).<\/span><\/li>\n<\/ul>\n<p><span style=\"font-family: tahoma, arial, helvetica, sans-serif;\"><img loading=\"lazy\" decoding=\"async\" class=\"alignnone size-full wp-image-2934\" src=\"https:\/\/praxilabs.com\/en\/blog\/wp-content\/uploads\/2024\/07\/Steps-of-Polyacrylamide-Gel-Electrophoresis-Simulation.jpg\" alt=\"\u00a0Steps of Polyacrylamide Gel Electrophoresis Simulation\" width=\"1200\" height=\"1050\" srcset=\"https:\/\/praxilabs.com\/en\/blog\/wp-content\/uploads\/2024\/07\/Steps-of-Polyacrylamide-Gel-Electrophoresis-Simulation.jpg 1200w, https:\/\/praxilabs.com\/en\/blog\/wp-content\/uploads\/2024\/07\/Steps-of-Polyacrylamide-Gel-Electrophoresis-Simulation-300x263.jpg 300w, https:\/\/praxilabs.com\/en\/blog\/wp-content\/uploads\/2024\/07\/Steps-of-Polyacrylamide-Gel-Electrophoresis-Simulation-1024x896.jpg 1024w, https:\/\/praxilabs.com\/en\/blog\/wp-content\/uploads\/2024\/07\/Steps-of-Polyacrylamide-Gel-Electrophoresis-Simulation-768x672.jpg 768w\" sizes=\"auto, (max-width: 1200px) 100vw, 1200px\" \/><\/span><\/p>\n<h3><span class=\"ez-toc-section\" id=\"_Steps_of_Polyacrylamide_Gel_Electrophoresis_Simulation\"><\/span><span style=\"font-family: tahoma, arial, helvetica, sans-serif;\"><span style=\"font-weight: 400; font-size: 14pt;\">\u00a0<\/span><span style=\"font-size: 14pt;\"><b>Steps of<\/b> <b>Polyacrylamide Gel Electrophoresis Simulation<\/b><\/span><\/span><span class=\"ez-toc-section-end\"><\/span><\/h3>\n<p><span style=\"font-weight: 400; font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\">By applying the following steps, you will perform the <a href=\"https:\/\/praxilabs.com\/en\/3d-simulations\/protein-electrophoresis-virtual-lab-simulation\">Polyacrylamide Gel Electrophoresis<\/a> simulation from PraxiLabs successfully:<\/span><\/p>\n<h4><span class=\"ez-toc-section\" id=\"First_Gel_and_buffer_preparation\"><\/span><span style=\"font-family: tahoma, arial, helvetica, sans-serif;\"><strong><span style=\"font-size: 14pt;\">First: Gel and buffer preparation<\/span><\/strong><\/span><span class=\"ez-toc-section-end\"><\/span><\/h4>\n<h4><span class=\"ez-toc-section\" id=\"Second_steps_of_gel_casting_and_sample_application\"><\/span><span style=\"font-family: tahoma, arial, helvetica, sans-serif;\"><strong><span style=\"font-size: 14pt;\">Second: steps of gel casting and sample application<\/span><\/strong><\/span><span class=\"ez-toc-section-end\"><\/span><\/h4>\n<p><span style=\"font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\"><span style=\"font-weight: 400;\">1. <\/span><b>Prepare the Casting Gel Cassettes:<\/b><\/span><\/p>\n<ul>\n<li><span style=\"font-weight: 400; font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\"> Clean the glass plates with distilled water, dry.<\/span><\/li>\n<li><span style=\"font-weight: 400; font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\"> Clamp two glass plates in the casting frames on the casting stands.<\/span><\/li>\n<li><span style=\"font-weight: 400; font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\"> Pipette distilled water and set aside until the\u00a0 gel is prepared to ensure no leakage.<\/span><\/li>\n<\/ul>\n<p><span style=\"font-family: tahoma, arial, helvetica, sans-serif;\"><b style=\"font-size: 14pt;\">2.Prepare the Separating Gel:<\/b><\/span><\/p>\n<p><span style=\"font-weight: 400; font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\"> To prepare a polyacrylamide 12%, 10 ml of separating gel:<\/span><\/p>\n<ul>\n<li><span style=\"font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\"> Remove D.W. by turning the cassette upside down on filter paper.<\/span><\/li>\n<li><span style=\"font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\">Pipette The appropriate amount of separating gel solution into the gap between the glass plates. (slowly, thoroughly), leave one 1 cm.<\/span><\/li>\n<li><span style=\"font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\"> To make the top of the separating gel be horizontal, fill in distilled water into the gap until it overflows.<\/span><\/li>\n<li><span style=\"font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\"> Wait for 20-30 min to let it gelate.<\/span><\/li>\n<li><span style=\"font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\"> Discard D.W. by turning the cassette upside down on filter paper.<\/span><\/li>\n<\/ul>\n<p><span style=\"font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\"><b>3.Prepare the Stacking Gel:<\/b><\/span><\/p>\n<p><span style=\"font-weight: 400; font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\"> To prepare a polyacrylamide 12%,use\u00a0 5 ml of stacking gel:<\/span><\/p>\n<ul>\n<li><span style=\"font-weight: 400; font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\"> Pipette the stacking gel until overflow.<\/span><\/li>\n<li><span style=\"font-weight: 400; font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\"> Insert the well-forming comb without trapping air under the teeth. Do not move it after insertion.<\/span><\/li>\n<li><span style=\"font-weight: 400; font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\"> Wait for 20-30 min to let it gelate. Make sure a complete gelation of the stacking gel.<\/span><\/li>\n<li><span style=\"font-weight: 400; font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\"> Take out the comb.<\/span><\/li>\n<li><span style=\"font-weight: 400; font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\"> Take the glass plates out of the casting frame and set them in the cell buffer unit.<\/span><\/li>\n<li><span style=\"font-weight: 400; font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\"> Pour the running buffer (electrophoresis buffer) into the inner chamber and keep pouring after overflow until the buffer surface reaches the mark on the outer chamber.<\/span><\/li>\n<\/ul>\n<p><span style=\"font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\"><b>4.Prepare samples:<\/b><\/span><\/p>\n<ul>\n<li><span style=\"font-weight: 400; font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\"> Add 5 ul sample in a tube.<\/span><\/li>\n<li><span style=\"font-weight: 400; font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\"> Add 3 ul loading buffer.<\/span><\/li>\n<li><span style=\"font-weight: 400; font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\"> Vortex for 10 seconds.<\/span><\/li>\n<li><span style=\"font-weight: 400; font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\"> Heat them at 95\u02daC for 3 min.<\/span><\/li>\n<\/ul>\n<p><span style=\"font-family: tahoma, arial, helvetica, sans-serif;\"><span style=\"font-weight: 400; font-size: 14pt;\"> Using the pipette, Load prepared samples into wells and make sure not<\/span><span style=\"font-weight: 400; font-size: 14pt;\">to overflow.<\/span><\/span><\/p>\n<ol>\n<li><span style=\"font-weight: 400; font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\"> Load 4 ul protein marker into the first lane.<\/span><\/li>\n<li><span style=\"font-weight: 400; font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\"> Cover the top and connect the electrodes.<\/span><\/li>\n<li><span style=\"font-weight: 400; font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\"> Set at 120 volt for one hour and run electrophoresis.<\/span><\/li>\n<li><span style=\"font-weight: 400; font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\"> Stop SDS-PAGE running when the lowermost sign of the protein marker almost reaches the foot line of the glass plate.<\/span><\/li>\n<\/ol>\n<h3><span class=\"ez-toc-section\" id=\"General_Guidelines_for_SDS-PAGE_SDS-_Polyacrylamide_Gel_Electrophoresis_Experiment\"><\/span><span style=\"font-family: tahoma, arial, helvetica, sans-serif;\"><b>General Guidelines for SDS-PAGE (SDS- Polyacrylamide Gel Electrophoresis) Experiment<\/b><\/span><span class=\"ez-toc-section-end\"><\/span><\/h3>\n<p>&nbsp;<\/p>\n<p><span style=\"font-family: tahoma, arial, helvetica, sans-serif;\"><img loading=\"lazy\" decoding=\"async\" class=\"alignnone size-full wp-image-2932\" src=\"https:\/\/praxilabs.com\/en\/blog\/wp-content\/uploads\/2024\/07\/General-Guidelines-for-SDS-PAGE-SDS-Polyacrylamide-Gel-Electrophoresis-Experiment.jpg\" alt=\"General Guidelines for SDS-PAGE (SDS- Polyacrylamide Gel Electrophoresis) Experiment\" width=\"1200\" height=\"628\" srcset=\"https:\/\/praxilabs.com\/en\/blog\/wp-content\/uploads\/2024\/07\/General-Guidelines-for-SDS-PAGE-SDS-Polyacrylamide-Gel-Electrophoresis-Experiment.jpg 1200w, https:\/\/praxilabs.com\/en\/blog\/wp-content\/uploads\/2024\/07\/General-Guidelines-for-SDS-PAGE-SDS-Polyacrylamide-Gel-Electrophoresis-Experiment-300x157.jpg 300w, https:\/\/praxilabs.com\/en\/blog\/wp-content\/uploads\/2024\/07\/General-Guidelines-for-SDS-PAGE-SDS-Polyacrylamide-Gel-Electrophoresis-Experiment-1024x536.jpg 1024w, https:\/\/praxilabs.com\/en\/blog\/wp-content\/uploads\/2024\/07\/General-Guidelines-for-SDS-PAGE-SDS-Polyacrylamide-Gel-Electrophoresis-Experiment-768x402.jpg 768w\" sizes=\"auto, (max-width: 1200px) 100vw, 1200px\" \/><\/span><\/p>\n<p style=\"text-align: center;\"><span style=\"font-size: 14pt; font-family: tahoma, arial, helvetica, sans-serif;\"><strong>Increase your Students\u2019 Learning Retention and Engagement!<\/strong><\/span><\/p>\n<p style=\"text-align: center;\"><span style=\"font-family: tahoma, arial, helvetica, sans-serif;\"><a href=\"https:\/\/praxilabs.com\/en\/request-free-demo\"><b>Request A Free Demo Now!<\/b><\/a><\/span><\/p>\n","protected":false},"excerpt":{"rendered":"<p>Polyacrylamide Gel Electrophoresis (PAGE) is a fundamental technique in the realm of molecular biology, enabling the separation and analysis of macromolecules such as DNA, RNA, and proteins based on their size and charge. As a cornerstone of scientific research and clinical diagnostics, understanding the intricacies of this method is essential. \u00a0In this blog, we&#8217;ll delve &hellip;<\/p>\n","protected":false},"author":8,"featured_media":4482,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"_lmt_disableupdate":"no","_lmt_disable":"no","footnotes":""},"categories":[6,1],"tags":[],"class_list":["post-2930","post","type-post","status-publish","format-standard","has-post-thumbnail","","category-biology","category-virtual-learning"],"modified_by":"Muhamed Elmesery","_links":{"self":[{"href":"https:\/\/praxilabs.com\/en\/blog\/wp-json\/wp\/v2\/posts\/2930","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/praxilabs.com\/en\/blog\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/praxilabs.com\/en\/blog\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/praxilabs.com\/en\/blog\/wp-json\/wp\/v2\/users\/8"}],"replies":[{"embeddable":true,"href":"https:\/\/praxilabs.com\/en\/blog\/wp-json\/wp\/v2\/comments?post=2930"}],"version-history":[{"count":11,"href":"https:\/\/praxilabs.com\/en\/blog\/wp-json\/wp\/v2\/posts\/2930\/revisions"}],"predecessor-version":[{"id":5169,"href":"https:\/\/praxilabs.com\/en\/blog\/wp-json\/wp\/v2\/posts\/2930\/revisions\/5169"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/praxilabs.com\/en\/blog\/wp-json\/wp\/v2\/media\/4482"}],"wp:attachment":[{"href":"https:\/\/praxilabs.com\/en\/blog\/wp-json\/wp\/v2\/media?parent=2930"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/praxilabs.com\/en\/blog\/wp-json\/wp\/v2\/categories?post=2930"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/praxilabs.com\/en\/blog\/wp-json\/wp\/v2\/tags?post=2930"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}