I found too many bands when ~30 microgram protein loaded in the gel from cancer cell lines, but I see very clean band in normal cells! Your precious comments will be appreciated! Thanks
I used to work with mouse and human breast cancer cell lines. I usually loaded 40 to 50 ug, and 100 for some specific proteins. Despite there were lots of bands on comassie blue I didn´t had that problem when reveling with specific antibodies. It actually depends of the antibody but in general It worked well.
What do you mean by "too many bands" - on coomassie/ponceau/after probing?
I've worked with breast and prostate cancer and normal cell lines. 12-25ug of protein has worked fine - even actin, most proteins of interest showed up fine. (Homemade gels, wet transfer at 70V-90min)
Dear Nawas,
I am talking about too many non specific bands when I probe with high molecular wt. antibody like 200KD. I can see the clean and specific band for small proteins e.g. Actin. I think you will understand!
Thank you for sharing your protein concentration and Transfer condition.
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