Protein Electrophoresis Reagent Selection and Preparation

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Overview

This section details how to select and prepare the reagents (protein standards, gels, and buffers) required for various SDS-PAGE applications. The types of gels and buffers selected should suit the size of the protein under investigation, the desired analysis technique, and the overall goals of the experiment.

Related Topics: Protein Standards, Polyacrylamide Gels, Buffer Systems and Gel Chemistries,and Handcasting Polyacrylamide Gels.


General Considerations

No particular gel type or buffer is useful for all proteins, and choosing the buffer systems and gel types that offer the highest resolution in the size range of interest may require some experimentation. In selecting reagents for SDS-PAGE, consider the following:

  • Protein standards — select protein standards that provide maximum resolution in the size range of interest and that offer compatibility and utility for downstream applications such as western blotting
  • Gel percentage — choose the percentage that offers resolution in the range of interest
  • Precast vs. handcast gels — precast gels offer greater convenience and superior quality control and reproducibility than handcast gels; handcast gels offer the possibility of customized percentages and gradients
  • Gel format — select mini- or midi-format gels when throughput is important or sample size is limited; select large-format gels for higher resolution. Select a comb type and gel thickness to accommodate the sample number and volume you are working with
  • Buffer system — choose the system that offers the best resolution and compatibility with the protein and application of interest
 

Related Content

 
Literature
Number Description Download
6040 Electrophoresis Guide, Interactive PDF, Rev C Click to download
 
 
LUSP8GD57 [x-forwarded-proto] = [http] [x-forwarded-port] = [80] [x-forwarded-for] = [69.166.8.45, 10.232.3.134] [seourl] = [/en-us/applications-technologies/applications-technologies/protein-electrophoresis-reagent-selection-preparation] [x-amzn-trace-id] = [Root=1-5c162fb5-351d9ff0f0627860fac335e0] [x-forwarded-server] = [lsds-prod-s.br.aws-livesite.io] [x-forwarded-host] = [www.bio-rad.com] [x-query-string] = [ID=LUSP8GD57] [host] = [10.232.0.21:1776] [x-request-uri] = [/en-us/applications-technologies/applications-technologies/protein-electrophoresis-reagent-selection-preparation] [connection] = [Keep-Alive] [cache-control] = [max-age=0] [accept-encoding] = [identity] [user-agent] = [Mozilla/5.0 (compatible; Googlebot/2.1; +http://www.google.com/bot.html)] AppTech/AppTechDetails pageStyleKey internet/solutions_sub applications-technologies/applications-technologies/protein-electrophoresis-reagent-selection-preparation LSR LUSP8GD57 Electrophoresis Reagent Selection and Preparation Protein Electrophoresis Reagent Selection and Preparation /webroot/web/html/lsr/solutions/technologies/protein_electrophoresis <p>This section details how to select and prepare the reagents (protein standards, gels, and buffers) required for various SDS-PAGE applications. The types of gels and buffers selected should suit the size of the protein under investigation, the desired analysis technique, and the overall goals of the experiment.</p> <p><strong>Related Topics:</strong> <a href="/evportal/destination/solutions?catID=LUSP9D7MU">Protein Standards</a>, <a href="/evportal/destination/solutions?catID=LUSPBRM5B">Polyacrylamide Gels</a>, <a href="/evportal/destination/solutions?catID=LUSPCXIDF">Buffer Systems and Gel Chemistries</a>,and <a href="/evportal/destination/solutions?catID=LUSPDTO62">Handcasting Polyacrylamide Gels</a>.</p> <h3 class="clear"><br /></h3> <h3 class="clear">General Considerations</h3> <p><img style="margin: 8px 0px 3px 0px;" src="/evportal/framework/skins/evolution/images/1x1grey.gif" border="0" alt="" width="620" height="1" /></p> <p>No particular gel type or buffer is useful for all proteins, and choosing the buffer systems and gel types that offer the highest resolution in the size range of interest may require some experimentation. In selecting reagents for SDS-PAGE, consider the following:</p> <ul> <li><a href="/evportal/destination/solutions?catID=LUSP9D7MU#applications">Protein standards</a> &mdash; select protein standards that provide maximum resolution in the size range of interest and that offer compatibility and utility for downstream applications such as western blotting</li> <li><a href="/evportal/destination/solutions?catID=LUSPBRM5B#percentage">Gel percentage</a> &mdash; choose the percentage that offers resolution in the range of interest</li> <li><a href="/evportal/destination/solutions?catID=LUSPBRM5B#precast_v_handcast">Precast vs. handcast gels</a> &mdash; precast gels offer greater convenience and superior quality control and reproducibility than handcast gels; handcast gels offer the possibility of customized percentages and gradients</li> <li><a href="/evportal/destination/solutions?catID=LUSPBRM5B#format">Gel format</a> &mdash; select mini- or midi-format gels when throughput is important or sample size is limited; select large-format gels for higher resolution. Select a comb type and gel thickness to accommodate the sample number and volume you are working with</li> <li><a href="/evportal/destination/solutions?catID=LUSPCXIDF#gel_buffer_chemistries_for_page">Buffer system</a> &mdash; choose the system that offers the best resolution and compatibility with the protein and application of interest</li> </ul> 6040 /templatedata/internet/documentation/data/LSR/Literature/6040.xml Life Science Research/Products/Electrophoresis and Blotting/Protein Electrophoresis and Blotting/Protein Standards/Recombinant Standards (Ladders)/Precision Plus Protein Unstained Standards ->MT::ab429c02-ab15-4967-a7b9-1f78850fc4d3##Life Science Research/Products/Electrophoresis and Blotting/Protein Electrophoresis and Blotting/Protein Standards/Recombinant Standards (Ladders)/Precision Plus Protein Prestained Standards ->MT::a7b0f9ce-e080-4b51-ab99-4cded66497c1##Life Science Research/Products/Electrophoresis and Blotting/Protein Electrophoresis and Blotting/Mini Format 1D-Electrophoresis Systems/Mini-PROTEAN Precast Gels/Mini-PROTEAN TGX Precast Gels ->MTS::KSHTFUE8Z## 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## Eddie C Selection & Preparation of Electrophoresis Gels 12/16/11 09:21 AM 12/16/21 09:22 AM 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
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