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Lidstrom: SDS-PAGE: Difference between revisions
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==Questions/Answers/Facts== | ==Questions/Answers/Facts== | ||
===What is the best type of control to test expression of your plasmid=== | |||
* If you are using SDS-PAGE to test for expression of a plasmid, include an empty vector version of the strain you are looking for. This is good because you can look for your band, which hopefully doesn't overlap with a band that already exists in the host strain. | |||
* This is also better than adding purified protein in a separate lane because the samples can run at different speeds. I'm not sure exactly why this happens, but here is an example: | |||
** [[image:150225_PAGE_JM.png|thumb|center|Do the bands run differently than the purified proteins just because a "smile" effect? Or is it something else like the density of the loading dye? Or something else?]] | |||
===How long are commercial PAGE gels good for after their expiration date?=== | ===How long are commercial PAGE gels good for after their expiration date?=== | ||
We have used gels that expired 2-3 years before their use date. Specifically they were BioRad TGX gels. -JM 3/2015 | We have used gels that expired 2-3 years before their use date. Specifically they were BioRad TGX gels. -JM 3/2015 | ||
They looked great! | They looked great! | ||
===How do you prepare soluble and insoluble fractions for SDS-PAGE?=== | ===How do you prepare soluble and insoluble fractions for SDS-PAGE?=== | ||
