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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.|
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Seeing a Shift in a control lane
I am doing EMSAs, and I am seeing a shift in a control lane where the cell have not been stimulated (Ie should not expect to see the wanted protiens in the nuclear extract.) the protein also should not be in a high concentration in the cytoplasm (NFKB)
The negative control (no nuclear extract added to the binding reaction) works fine (no shift-->no protien bound)
does this mean that I am having nonspecific protiens binding to my DNA?
Also, this control lane (no stimulation to cells) has a shift with the same size as the experimental lanes... so I don't even know if the experimental lanes are true based on this.
do you think its a problem with the binding reaction, or Nuclear extraction methods?
|control , lane , shift|
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