B-actin as western blot -control
Posted 11 July 2010 - 05:37 PM
Posted 13 July 2010 - 01:30 AM
Posted 15 July 2010 - 04:41 PM
I think there unequal loading of proteins thats why beta actin not exact in each sample.
may be mistake in calculating protein conc . or in protein assay. check out this two possibilities..
Posted 19 July 2010 - 09:21 AM
Although we tend to think the so called "house-keeping" genes should not change with conditions, they do. Perhaps b-actin does change in your tumor samples, if you are sure that you have loaded equal amount of protein...I would try it again, just in case i have done my maths wrong, or loaded more/less volume of the samples, or some how i haven't got an equal transfer...and if the problem persists, you might want to explore other house-keeping genes as loading control...
Yes equal amounts of protein was loaded, however where u require 7 microlitres for tumor, normal sample requires about 30 microlitres. does it effect when the wells are loaded almost full to its capacity? Will it affect the resolution and weaker bands?
Posted 19 July 2010 - 09:48 AM
genius does what it must
i do what i get paid to do