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Tips for Successful SDS-PAGE Gel Electrophoresis

A list of tips for getting beautiful SDS-PAGE Gels.

Hints for SDS-PAGE Gel Electrophoresis Table of Contents

SDS-PAGE Tips
Background
SDS-PAGE Videos
Other SDS-PAGE Protocols
Troubleshooting SDS-PAGE Gels
Discussion Topics for Agarose Gel Electrophoresis
Get Help on your Agarose Gel Problem

Tips on How to get Beautiful SDS-PAGE Gels

Always use the highest quality acrylamide and bis for your gels.
Loading buffer is very important for the quality of the running of your samples. Always make your own or purchase the best. A great recipe is Laemmeli Sample Buffer. If your samples are puffing out of the loading lanes, you need more sucrose to weight your samples down in your sample buffer (either that or mix your sample buffer well before using it the sucrose may be at the bottom).
When preparing samples the protein samples should be mixed by vortexing before and after the heating step for best resolution.
Samples should not be boiled at 100 degrees celsius! Boiling at high temperatures can lead to high molecular weight aggregates. Reference: Thermal aggregation of SARS-CoV membrane protein, Lee et al., 2005. Many find it important to avoid boiling our samples prior to SDS-PAGE for membrane proteins involved in lipid synthesis. Boiling can lead to aggregation of proteins. You can just heat your sample to 65 degrees in SDS sample buffer.
To avoid gel edge effects you can add 1x sample buffer to your unused well lanes.
If you are running your gels in the cold room or in a cold environment, look into using lithium dodecyl sulfate (LiDS) instead of SDS as LiDS does not precipitate at lower temperatures.
Most researchers use constant power conditions when running gel electrophoresis however it was determined that only constant voltage gives constant protein mobility during gel electrophoresis.

SDS-PAGE Gel Electrophoresis Forum Topics

western blot Hi, i'm doing my research on plant model. So far we did not use any equal load control, the actin Ab we have in our laboratory doesn' work. So, i'm...
proteins not running Hello. I'm having a problem with proteins getting "stuck" at the top of the gel and not running correctly. This is a new problem in the past month....
Cracked wells in SDS page hello everybody i wanted to know if uneven or distorted wells could cause the bands to be distorted?:wiggle:
PAGE storage and tubulin antibodies Hello, 1. Once the PAGE is prepared, how long can it be stored and under what conditions? For instance, if I prepare the gel on Friday afternoon,...
sds-page sample contains enzyme but in page bands are not clear
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SDS-PAGE Electrophoresis Videos

Demystifying SDS-Page Video

An instructional video aimed at teaching the process of SDS-PAGE to college students with basic biochemical understandings. www.sdspage.net From Youtube user mattbrewbaker.

Added by: kiki06
Tags: sds-page gel electrophoresis
Date: 2008-04-28

SDS-PAGE Gel Electrophoresis Video