Introduction to Protein Electrophoresis

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Photo of protein electrophoresis gel and gel box

Overview

Gel electrophoresis can be used to separate proteins for both analysis and purification. This section provides a brief overview of the theory and workflow of protein electrophoresis.

Related Topics: Protein Transfer, Immunodetection, and Imaging.

 

How Protein Electrophoresis Works

The rate at which proteins move in an electrical field (migration rate, in units of cm2 V-1 sec-1) is governed by a complex relationship between the physical characteristics of both the electrophoresis system and the proteins. The strength of the electric field, the properties of the electrophoretic medium (usually a polyacrylamide gel), the temperature of the system, and the pH, ion type, and concentration of the buffer all play roles along with the size, shape, and charge of the proteins (Garfin 1990). Proteins come in a wide range of size and shapes and have charges determined by the dissociation constants of their constituent amino acids. As a result, proteins have characteristic migration rates that can be exploited for separation purposes. Protein electrophoresis can be performed in liquid or gel-based media and can also be used to move proteins from one medium to another (for example, in blotting applications).

Diagram of protein migration in electrophoresis

Movement of proteins during electrophoresis.

Protein electrophoresis can be used in a variety of applications including protein purification or purity determination (for example, at various stages during a chromatographic purification), to determine size, isoelectric point (pI), and enzymatic activity, or to provide data on the regulation of protein expression (Dunn 1993). In fact, a number of different techniques can be grouped under the term protein electrophoresis including gel electrophoresis, isoelectric focusing (IEF), and two-dimensional (2-D) electrophoresis.

 

General Considerations in Protein Electrophoresis

A electrophoresis workflow involves the selection of the appropriate method, instrumentation, and reagents for the intended experimental goal.

Methods include the following:

Once proteins have been separated, they can be utilized for a number of downstream applications including enzymatic assays, further purification, transfer to a membrane for immunological detection (immunoblotting or western blotting), and elution and digestion for mass spectrometric analysis.

 

Protein Electrophoresis Workflow

Diagram of protein electrophoresis workflow Method Selection Sample Preparation Reagent Selection and Preparation Performing Electrophoresis Protein Detection and Analysis

A typical protein electrophoresis workflow.

Bio-Rad offers a wide range of protein gels in optimized formulations for SDS-PAGE, native PAGE, and specialized gel chemistries.

Learn more »
Bio-Rad Protein Gels
 

References for Protein Electrophoresis

Dunn MJ (1993). Gel electrophoresis of proteins (Abingdon Oxford, United Kingdom: BIOS Scientific Publishers Ltd.).

Garfin DE (1990). One-dimensional gel electrophoresis. Methods Enzymol 182, 425–441.

 

Related Content

 
Literature
Number Description Download
6040 Electrophoresis Guide, Interactive PDF, Rev C Click to download
2651 2-D Electrophoresis Workflow How-To Guide, Rev F Click to download
2895 Protein Blotting Guide, Ver C Click to download
 
 
LUSOVO47B [sec-fetch-mode] = [navigate] [referer] = [https://www.google.com/] [sec-fetch-site] = [cross-site] [x-forwarded-proto] = [https] [accept-language] = [en-US,en;q=0.9] [x-forwarded-port] = [443] [x-forwarded-for] = [157.50.153.14, 10.232.19.132] [sec-fetch-user] = [?1] [accept] = [text/html,application/xhtml+xml,application/xml;q=0.9,image/avif,image/webp,image/apng,*/*;q=0.8,application/signed-exchange;v=b3;q=0.9] [seourl] = [/en-us/applications-technologies/introduction-protein-electrophoresis] [x-amzn-trace-id] = [Root=1-5f72b5b9-295f3164e1c41946b32cd13e] [x-forwarded-server] = [lsds-prod-s.br.aws-livesite.io] [x-forwarded-host] = [www.bio-rad.com] [x-query-string] = [ID=LUSOVO47B] [host] = [10.232.0.21:1776] [upgrade-insecure-requests] = [1] [x-request-uri] = [/en-us/applications-technologies/introduction-protein-electrophoresis] [connection] = [Keep-Alive] [accept-encoding] = [gzip, deflate, br] [user-agent] = [Mozilla/5.0 (Windows NT 6.1) AppleWebKit/537.36 (KHTML, like Gecko) Chrome/85.0.4183.121 Safari/537.36] [sec-fetch-dest] = [document] AppTech/AppTechDetails pageStyleKey internet/solutions_sub applications-technologies/introduction-protein-electrophoresis LSR LUSOVO47B Protein Electrophoresis Introduction to Protein Electrophoresis /webroot/web/html/lsr/solutions/technologies/protein_electrophoresis /webroot/web/images/lsr/solutions/technologies/protein_electrophoresis_blotting_and_imaging/protein_electrophoresis/technology_detail/protein-electrophoresis-feature.jpg /webroot/web/images/lsr/solutions/technologies/protein_electrophoresis_blotting_and_imaging/technology_thumb/cat_pet1_protein_electro_icon.jpg Photo of protein electrophoresis gel and gel box <p>Gel electrophoresis can be used to separate proteins for both analysis and purification. This section provides a brief overview of the theory and workflow of protein electrophoresis.</p> <p><strong>Related Topics:</strong> <a href="/evportal/destination/solutions?catID=LUSPTIMNI">Protein Transfer</a>, <a href="/evportal/destination/solutions?catID=LUSQ4HE8Z">Immunodetection</a>, and <a href="/evportal/destination/solutions?catID=LUSQ8NC4S">Imaging</a>.</p> How Protein Electrophoresis Works <p>The rate at which proteins move in an electrical field (migration rate, in units of cm<sup>2</sup> V<sup>-1</sup> sec<sup>-1</sup>) is governed by a complex relationship between the physical characteristics of both the electrophoresis system and the proteins. The strength of the electric field, the properties of the electrophoretic medium (usually a polyacrylamide gel), the temperature of the system, and the pH, ion type, and concentration of the buffer all play roles along with the size, shape, and charge of the proteins (Garfin 1990). Proteins come in a wide range of size and shapes and have charges determined by the dissociation constants of their constituent amino acids. As a result, proteins have characteristic migration rates that can be exploited for separation purposes. Protein electrophoresis can be performed in liquid or gel-based media and can also be used to move proteins from one medium to another (for example, in <a href="/evportal/destination/solutions?catID=LUSPPAKG4">blotting</a> applications).</p> <p><img src="/webroot/web/images/lsr/solutions/technologies/protein_electrophoresis_blotting_and_imaging/protein_electrophoresis/technology_detail/protein-migration-protein-electrophoresis.jpg" alt="Diagram of protein migration in electrophoresis" width="279" height="298" /></p> <p class="caption"><strong>Movement of proteins during electrophoresis</strong>.</p> <p>Protein electrophoresis can be used in a variety of applications including protein purification or purity determination (for example, at various stages during a chromatographic purification), to determine size, isoelectric point (pI), and enzymatic activity, or to provide data on the regulation of protein expression (Dunn 1993). In fact, a number of different techniques can be grouped under the term protein electrophoresis including gel electrophoresis, <a href="/evportal/destination/solutions?catID=LUSQLK2B7">isoelectric focusing</a> (IEF), and <a href="/evportal/destination/solutions?catID=LUSQF04EH">two-dimensional (2-D) electrophoresis</a>.</p> <div class="top"><a href="#helptop">Back to Top</a></div> General Considerations in Protein Electrophoresis <p>A <a href="#protein_electrophoresis_workflow">electrophoresis workflow</a> involves the selection of the appropriate method, instrumentation, and reagents for the intended experimental goal.</p> <p>Methods include the following:</p> <ul> <li>Various types of polyacrylamide gel electrophoresis (PAGE) <ul> <li><a href="/evportal/destination/solutions?catID=LUSOW4GRI#dn_page">Discontinuous Native PAGE</a></li> <li><a href="/evportal/destination/solutions?catID=LUSOW4GRI#sds_page">SDS-PAGE</a></li> <li><a href="/evportal/destination/solutions?catID=LUSOW4GRI#bn_page">Blue Native PAGE</a></li> <li><a href="/evportal/destination/solutions?catID=LUSOW4GRI#zymogram_page">Zymogram PAGE</a></li> <li><a href="/evportal/destination/solutions?catID=LUSOW4GRI#ief">Isoelectric Focusing (IEF)</a></li> <li><a href="/evportal/destination/solutions?catID=LUSOW4GRI#two-d_electrophoresis">2-D Electrophoresis</a></li> <li><a href="/en-us/category/experion-automated-electrophoresis-system?ID=9026a8bd-2711-496d-b2d0-5f7a056cc26f">Experion&trade; Automated Electrophoresis System</a></li> </ul> </li> <li>Instrumentation needed for protein electrophoresis includes: <ul> <ul> <li><a href="/evportal/destination/solutions?catID=LUSOZFBEB#electrophoresis_cells">Electrophoresis cells</a></li> <li><a href="/evportal/destination/solutions?catID=LUSOZFBEB#power_supplies_for_PAGE_apps">Power supplies</a></li> </ul> </ul> <p>&nbsp;</p> </li> </ul> <p>Once proteins have been separated, they can be utilized for a number of downstream applications including enzymatic assays, further purification, transfer to a membrane for immunological detection (immunoblotting or <a href="/evportal/destination/solutions?catID=LUSPPAKG4">western blotting</a>), and elution and digestion for mass spectrometric analysis.</p> <div class="top"><a href="#helptop">Back to Top</a></div> Protein Electrophoresis Workflow <p><a name="protein_electrophoresis_workflow"></a></p> <p><img usemap="#Map" src="/webroot/web/images/lsr/solutions/technologies/protein_electrophoresis_blotting_and_imaging/protein_electrophoresis/technology_detail/protein-electrophoresis-workflow-protein-electrophoresis.jpg" border="0" alt="Diagram of protein electrophoresis workflow" width="531" height="710" /> <map name="Map"> <area shape="rect" coords="55,8,230,38" href="/evportal/destination/solutions?catID=LUSOW4GRI" alt="Method Selection" /> <area shape="rect" coords="55,151,225,176" href="/evportal/destination/solutions?catID=LUSP108WI" alt="Sample Preparation" /> <area shape="rect" coords="53,305,227,335" href="/evportal/destination/solutions?catID=LUSP8GD57" alt="Reagent Selection and Preparation" /> <area shape="rect" coords="54,469,227,495" href="/evportal/destination/solutions?catID=LUSPFBNEL" alt="Performing Electrophoresis" /> <area shape="rect" coords="54,624,226,651" href="/evportal/destination/solutions?catID=LUSPLU15" alt="Protein Detection and Analysis" /> </map></p> <p class="caption"><strong>A typical protein electrophoresis workflow.</strong></p> <div class="bannerAT"> <div class="bannerText ddpcrText" style="width: 404px !important;"> <h2 class="banner_header">Find the right gel for your application.</h2> <p>Bio-Rad offers a wide range of protein gels in optimized formulations for SDS-PAGE, native PAGE, and specialized gel chemistries.</p> <a class="linkgeneration" href="/_locale/category/protein-gels?at_banner=Protein Electrophoresis&amp;at_banner_rev=Protein Electrophoresis&amp;at_banner_e=c">Learn more &raquo;</a></div> <img src="/webroot/web/images/lsr/global/english/solutions/bio-rad-protein-gels.jpg" alt="Bio-Rad Protein Gels" width="615" height="120" /></div> <div class="top"><a href="#helptop">Back to Top</a></div> <script src="/webroot/web/js/countrySpecific-min.js" type="text/javascript"></script> <script type="text/javascript">// <![CDATA[ $(document).ready(function(){ setSterlingUrlsToHtmlHrefVariables(); $('a.linkgeneration').each(function(){ $(this).attr('href', $(this).attr('href').replace('_locale', languageCode + '-' + countryCode).replace('_verticalUrl', currentVerticalUrlTitle).replace('_defaultVerticalUrl', defaultVerticalUrlTitle).replace('_feedbackCMSID',feedbackCMSID)); }); }); // ]]></script> References for Protein Electrophoresis <p>Dunn MJ (1993). Gel electrophoresis of proteins (Abingdon Oxford, United Kingdom: BIOS Scientific Publishers Ltd.).<strong></strong></p> <p>Garfin DE (1990). One-dimensional gel electrophoresis. Methods Enzymol 182, 425&ndash;441.</p> 6040 /templatedata/internet/documentation/data/LSR/Literature/6040.xml 2651 /templatedata/internet/documentation/data/LSR/Literature/2651_0910140934647.xml 2895 Life Science Research/Products/Electrophoresis and Blotting/Protein Electrophoresis and Blotting/Mini Format 1D-Electrophoresis Systems/Mini-PROTEAN Tetra Cell Systems ->MT::5cf78e19-7ed5-4373-a988-3e62456a488e##Life Science Research/Products/Electrophoresis and Blotting/Protein Electrophoresis and Blotting/Midi Format 1D-Electrophoresis Systems/Criterion Cell Systems ->MT::b94e30f5-0f2a-452d-a3a0-825e8205b476##Life Science Research/Products/Electrophoresis and Blotting/Protein Electrophoresis and Blotting/Large Format 1D-Electrophoresis Systems/PROTEAN II xi Cells ->MT::030e1125-d5c2-4f5a-8d9f-a004d783b845##Life Science Research/Products/Electrophoresis and Blotting/Protein Electrophoresis and Blotting/2D-Electrophoresis/1st Dimension- Isoelectric Focusing (IEF)/PROTEAN IEF Cell ->MT::92682b19-051c-464f-8a73-97cd256b225f##Life Science Research/Products/Electrophoresis and Blotting/Experion Automated Electrophoresis System/Experion Automated Electrophoresis Station ->MT::e88b061b-37dc-4610-b4b7-0ef925f622f9## Karen Moss Introduction to Protein Electrophoresis Protocols, video tutorials, and selection guides to help you at every step of your electrophoresis experiments. protein electrophoresis, electrophorosis, elctrophoresis, electropheresis, protein purification, protein analysis 12/05/11 04:56 PM 12/05/21 05:12 PM AE,AI,AL,AM,AR,AT,AU,AZ,BA,BD,BE,BF,BG,BH,BN,BO,BR,BW,CA,CH,CL,CM,CN,CO,CR,CY,CZ,DE,DK,DO,DZ,EC,EE,EG,EH,ER,ES,ET,FI,FM,FO,FR,GA,GE,GF,GH,GP,GR,GT,GU,HK,HN,HR,HT,HU,ID,IE,IL,IN,IS,IT,JM,JO,JP,KE,KH,KR,KW,KZ,LB,LI,LK,LT,LU,LV,MA,MD,MG,MK,ML,MO,MQ,MS,MT,MU,MX,MY,NG,NI,NL,NO,NP,NZ,OM,PA,PE,PF,PG,PH,PK,PL,PR,PS,PT,PW,PY,QA,RO,RS,RU,SA,SB,SE,SG,SI,SK,SN,ST,SV,TG,TH,TN,TO,TR,TT,TW,TZ,UA,UG,UK,US,UY,UZ,VA,VE,VU,XK,YE,ZA en LSR /LSR/Technologies/Protein_Electrophoresis N 0 Life Science Research/Solutions/Technologies/Western Blotting ->MTS::LUSPPAKG4## Life Science Research/Solutions/Technologies/2-D Electrophoresis ->MTS::LUSQG6LPT## Life Science Research/Solutions/Applications/Protein Separation and Analysis ->MTS::LUSOU3F7Q##Life Science Research/Solutions/Applications/2-D Electrophoresis and Analysis ->MTS::LUSQF04EH##Life Science Research/Solutions/Applications/Automated Electrophoresis and Analysis ->MTS::LUSQXB15##Life Science Research/Solutions/Technologies/Automated Electrophoresis System -Experion- ->MTS::LUSR04KG4## Life Science Research/Solutions/Technologies/Western Blotting ->MTS::LUSPPAKG4## Life Science Research/Solutions/Technologies/2-D Electrophoresis ->MTS::LUSQG6LPT## Life Science Research/Solutions/Applications/Protein Separation and Analysis ->MTS::LUSOU3F7Q##Life Science Research/Solutions/Applications/2-D Electrophoresis and Analysis ->MTS::LUSQF04EH##Life Science Research/Solutions/Technologies/Automated Electrophoresis System -Experion- ->MTS::LUSR04KG4## 0
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