Electrophoretic Separation of Proteins
Bulbul Chakavarti, Deb Chakavarti
Proteomic Center, Keck Graduate Institute of Applied Life Sciences
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0:00 Title: Electrophoretic0 0:21 Subscription Lock21 0:45 Introduction45 1:06 Pour the Separating Gel66 2:44 Pour The Stacking Gel164 3:55 Prepare The Sample and Load the Gel235 6:24 Run the Gel384 7:59 Conclusion479
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Electrophoresis is used to separate complex mixtures of proteins (e.g., from cells, subcellular fractions, column fractions, or immunoprecipitates), to investigate subunit compositions, and to verify homogeneity of protein samples. It can also serve to purify proteins for use in further applications. In polyacrylamide gel electrophoresis, proteins migrate in response to an electrical field through pores in a polyacrylamide gel matrix; pore size decreases with increasing acrylamide concentration. The combination of pore size and protein charge, size, and shape determines the migration rate of the protein. In this unit, the standard Laemmli method is described for discontinuous gel electrophoresis under denaturing conditions, i.e., in the presence of sodium dodecyl sulfate (SDS).
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Chakavarti B, Chakavarti D (2008). Electrophoretic Separation of Proteins. JoVE. 16. http://www.jove.com/index/details.stp?id=758, doi: 10.3791/758
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Allowed tags: i, b, u, sup, sub 
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| | 06/15/2008 1:11:45 PM
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swetha responded with a statement of type: Neutral
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Hi nice demo but I did not understand why the presenters kept on showing an agarose gel electrophoresed DNA gel in between as a representation instead of a protein gel. |
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SYPROorange and SYPROruby staining - two fluorescent-based protein detection methods - can appear like ethidium bromide stained DNA gels. So those gels you are speaking of may actually be protein gels. |
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| | 06/16/2008 9:52:09 AM
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swetha responded with a statement of type: Neutral
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Yeah possible, but then why do we see only a single band.I guess SYPRO detection methods would detect the entire protein panel and not specific bands. unless otherwise they are detecting some specific enzyme activity using flourescent detection methods. |
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| | 06/17/2008 2:08:43 AM
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Bulbul Chakravarti responded with a statement of type: Neutral
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Please not that we showed polyacrylamide gel electrophoresis and not agarose gel electrophoresis.
Thanks.
Bulbul Chakravarti |
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| | 10/10/2008 3:35:20 AM
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karinahui responded with a statement of type: Neutral
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why cannot i see the video...? pls help me.. |
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| | 10/10/2008 4:50:52 AM
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karinahui responded with a statement of type: Neutral
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i can see it now..thanks... |
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| | 03/15/2009 6:30:14 AM
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lymichong responded with a statement of type: Neutral
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my network is poor, I can not enjoy the video |
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| | 03/22/2009 10:26:19 PM
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nikitab responded with a statement of type: Neutral
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Hello,
Please shoot me an email at nikitab@jove.com and we'll figure something out for you.
Cheers,
Nikita |
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Mailer.URL: http://www.jove.com/index/details.stp?id=758 |
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