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Protein Science Post Questions and Discuss Protein Structure and Function, Protein Localization etc.. Please do not post Proteomic questions here, there is a Proteomic Forum below.

Protein Science
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Protein Pictures
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Protein Pictures
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by admin
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Protein Pictures
16 photos
9 comments


SDS-PAGE Gel Thickness vs Loading

Protein Science

Post Questions and Discuss Protein Structure and Function, Protein Localization etc.. Please do not post Proteomic questions here, there is a Proteomic Forum below.



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Old 11-07-2007, 02:13 PM
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Question SDS-PAGE Gel Thickness vs Loading

Hello All,
I am new tho theis forum but I am hopeful someone out there can help me. I'm currently in the process of changing our gel systems to hopefully a more reliable pre-cast gel. We currently pour our own gels and they measure 0.75 mm thick. We are changing to a pre cast gel that is 1.0mm thick.

The question I have is, do I need to increase the volume of the load, increase the protein load, both, neither ?

On our current system we load 1.6µg protein. Should the protein load be increased to 2.13µg to compensate for the thicker gel? Our data seems to support that an increased load will yeild "equivalent" results. We scan to determine shoulder peak height percent as part of our release criteria. Our increased load for the thicker gel was calculated by simply determining that the gel is 33% thicker, therfore the protein load should be 33% more. I'm not sure if this is accurate and that is why I am looking for help. I have more experience scaling up HPLC methods and columns but I have very little knowledge when it comes to changing SDS-PAGE coomassie systems.

Thanks for your help.
PPMQC
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