double bands on p-p70s6k WB - (Sep/06/2019 )
Ran a WB for p-p70s6k with SC Ab(9208), got a 85kDa bands as expected. but it showed double bands of 70kDa and the 70KdA bands look lower than 70kDa. Any help would be appreciated. Thank you.
What were your conditions for blotting and antibody detection, and what steps have you tried to eliminate non-specific bands? What does the literature, including the product website say about this antibody?
If you are using this antibody: https://www.cellsignal.com/products/primary-antibodies/phospho-p70-s6-kinase-ser371-antibody/9208 Note the sizes mentioned...
They may be artifacts caused by aged 2-mercaptoethanol (some have identified them as keratin from dust). You can test this by running a sample without protein, but everything else, including the buffer in which your sample is solubilized.
These artifacts usually show up as bands lower than bsa (around 40-50 kDa).
Thank you for your suggestions. I found a paper published on hematology. The s6k western also showed two bands.They didn't explain this.