SDS PAGE Problem - SDS page (Jul/07/2008 )
im running my sds page with soluble proteins(centrifuge from broth and dissolve in 0.9% NaCl) from Vibrio parahaemolyticus.. the problem is that there is no proteins bands but the molecular weight marker was visible, why is this so?
ive boil the sample for 5 mins with sds reducing buffer from bio rad , i used 10% polyacrylamide gel both in resolving and stacking gel... i also used biorad prepared buffer for sds page.
does the stacking and the resolving gel should have different concentrations?
thanks in advance
Have you tried increasing the amount of protein loaded on your gel?
10% gels should be okay for proteins ranging from 20 to 250 kDa.. so you should see something.
. you can check the concentration of your protein extracts before loading
. you can do 2 gels in same time. Color one with coomassie blue and tranfer the second only if you see bands in the stained gel
. you have not enough proteins to see on ponceau staining but maybe sufficent for antibody detection