Refer to the picture attached, it shows some of the lanes doesnt show a nice band. May I know any possible reasons or causes that make this happen?
The smearing in lanes 2 and 3 are either from too much protein, too much 2nd Ab or exposing to the ECL reagents for too long. Less is more.
Good luck!
It looks like these effects might be caused at the step of ECL development of your membrane (for example, touching the spot of the bands after ECL development).
I get smears like that as a result of insufficient blocking, then some Ab (1st, 2nd or both) attach directly to the membrane.
Thanks for the answers provided.. Perhaps I should change my question.
By comparing figure A (WB) and figure B (WB2), why the lanes 1 to 9 in figure A showing some smearing or not so good looking band as compare to figure B?
Figure B is a different gel, I suppose? Did you use precast or selfcast gels? Inhomogeneities of the gel could have also affected the bands in figure A in terms of protein migration and/or transfer (eg the bend in lane 8).
Hi Alexander, figure B is a different gel.. but I used the same receipt for both gels and run them at the same time and same voltage.. the antibodies I used for both of them were same
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