I ran a 15ug western blot using protein from treated mammalian cells. The top bands are Ligase 4 and the bottom bands are B-Actin. I am not sure why there is smearing in the Ligase 4 bands but not the B-Actin bands. The Ligase 4 antibody has worked previously, but I am not sure why it isn't working anymore.
Domenico Sanfelice
It looks to me like there is a problem with the samples rather then the antibody, in fact line 4 and 11 are fine. Did u load the same amount in all lanes/are they lysed in the same way? I get bands like that when the gel is overloaded or when the sample is very gloopy you should improve that!
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Melissa Chavez
I loaded the concentration of 15ug per well and they were lysed the same way. And thank you for the comment
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