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Electrophoretic Mobility Shift Assay Forum EMSA Forum. Discuss the Electrophoretic Mobility Shift Assay protocols or technique and the assessment of DNA and RNA binding proteins here. Also post questions and troubleshooting about the shift assay.


Does anybody knows how to interpret Non radioactive EMSA

Does anybody knows how to interpret Non radioactive EMSA - Electrophoretic Mobility Shift Assay Forum

Does anybody knows how to interpret Non radioactive EMSA - EMSA Forum. Discuss the Electrophoretic Mobility Shift Assay protocols or technique and the assessment of DNA and RNA binding proteins here. Also post questions and troubleshooting about the shift assay.


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Old 03-07-2012, 07:59 AM
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Default Does anybody knows how to interpret Non radioactive EMSA



Hi all,

I am attaching my EMSA result I know it is not perfect because it needs more optimization, however, I realized that I have very small shift in lane 5. Also,
I realized that when there is a shift, the shifted band becomes more fainter in color compared with the free RNA band, does anybody has explanation for this?


from right to left:

1- free RNA
2-biotinylated rna+ protein 100nm
3-biot. rna + protein 200nm
4-biot. rna+protein 400nm
5-biot. rna+protein 600 nm
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Old 03-15-2012, 04:07 PM
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Default Re: Does anybody knows how to interpret Non radioactive EMSA

Quote:
Originally Posted by DNAIA View Post
Hi all,

I am attaching my EMSA result I know it is not perfect because it needs more optimization, however, I realized that I have very small shift in lane 5. Also,
I realized that when there is a shift, the shifted band becomes more fainter in color compared with the free RNA band, does anybody has explanation for this?


from right to left:

1- free RNA
2-biotinylated rna+ protein 100nm
3-biot. rna + protein 200nm
4-biot. rna+protein 400nm
5-biot. rna+protein 600 nm
folder
Attached Files
File Type: zip emsa 2.zip (41.5 KB, 5 views)
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Old 03-31-2012, 09:51 PM
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Default Re: Does anybody knows how to interpret Non radioactive EMSA

Which way round is your film?
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Old 05-11-2012, 10:55 AM
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Default Re: Does anybody knows how to interpret Non radioactive EMSA

Form its look like RNA-protein complex migrates faster than the free probe. It means that RNA in complex with protein has quiet diferent structure than alone. If the orientation of the gel is correct than my explanation is like above.

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