I'm using the MDA-MB-231 cell line and looking into the expression of certain proteins. Is it possible to quantify them either in the nucleus or in the cytoplasm in order to compare between them?
You can try by IFA assay and confocal microscopy using software quantification of the same microscope, quantifies the intensity of the signal recognition in the nucleus, marking the area of the nucleus and compare it with the intensity of the total cell, now subtracting the signal from the nucleus, and so you get the difference. You can even get statistical data if you make the right reps. Greetings!
Thank you so much Jonathan.
I am not sure if the software of our microscope can mark the area of the nucleus and only quantify the intensity of the signal recognition in it but I'll absolutely try it. Thank you again.
I suggest you to try western blotting by extracting both nuclear and cytoplasmic fractions.If you are interested in phosphorylation status, WB will also helps. Here you can find an example of article mentioning comparison of nuclear and cytoplasmic status of a certain protein complex.
http://hmg.oxfordjournals.org/content/17/19/2934.short
There are usefull hommade protocols for nuclear and cytoplasmic fraction isolation, or you can use commercial kits such as Thermo or Millipore.
Hope this helps.
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