Western blot gel running problem - Gel Electrophoresis (Aug/08/2006 )
I'm using Precise Protein Gels 4-20% , Tris-HEPES-SDS running buffer , the problem is that I got diffused running leans in all of the wells even the blanks (loading buffer only), the way I always run my gels I get sharp straight line, but not this time I wonder whether it is a problem of the electrophoresis system???
This is the first time this problem had happen to me, I do not know what went wrong and I hope to find an answer from you...
It happended to me, it was a problem with SDS.
When I arrived in this new lab, I used the SDS solution that was bought here.
My bands were awfull, large, and not linear.
It took me sometime to realize it could be the SDS. I found some powder in a closet, prepared my own solution, and now my bands are perfect.
I think I realized what the problem was; I prepared the running buffer the weekend before I do the WB.
I don't think this is the problem. I prepare my running buffer several days before without any problem.
I prepared my running buffer as 10x in Jan and am still using it. It works just fine.
Can anyone tell me how to calculate the (micro molar)/molar wt. of a protein given the protein's concentration in mg/ml (e,g. 1mg/ml) and the wt in Kda?
you can find what you need here; it is a neat tool I got from fred
the molecular weight in kDa it's the number of gram in one mole.
then a 14 kDa protein is 14000g/mol