Hi
I am working on lair2 and it's interaction with C1q to understand who that affect lupus.
We used cell line to produce lair2 . than we transfected it with c1q.
and now we are doing western blot , to detect the two proteins, the first time we incubate them with c1q antibody . and we see the two proteins . after that we used the same membrane to detect lair2 , we wash the membrane and incubate it with lair 2 antibody and we see the protein .
we used 2 negative controls , in the first detection we saw two faint bands in them . in the second one we didn't see them.
I don't know what's the problem , and how to analyze it?
I had a similar problem, the second detection was very weak, so I tried to divided my samples by half (repeated loading into the gel in the same order), and then after the transfer I cut the membrane and each half was detected by different antibodies, it worked every time.
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