Lanes 1 to 8 from left to right represent protein from chondrocyte cell line. I ip'd with 5ug of antibody both igg and sox9 from 450ug of total protein. I have no input control as I ran out of protein. I've also attached the coomassie blue staining that I did POST transfer. Thanks again.
1: IP with sox9
2: IP with igg
3: IP with sox9
4: IP with igg:
5: IP with sox9
6: IP with igg
7: IP with sox9
8: IP with igg
Duco Kramer
Either you have overloaded the gel (to much total protein per lane) or the salt concetration of the samples is to high.
The first: try to load half or 25% of the sample.
2nd: after the final washing of the IP-samples take the beads up in 25 ul 1x Laemmly (or whatever running buffer you prefer).
Good luck
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