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double bands on p-p70s6k WB

western blotting

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#1 ylu

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Posted 06 September 2019 - 05:23 AM

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.

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  • 2019-09-05 huh Pp70S69208 s.jpg


#2 bob1

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Posted 06 September 2019 - 04:09 PM

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.cellsign...1-antibody/9208 Note the sizes mentioned...



#3 mdfenko

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Posted 08 September 2019 - 07:23 PM

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).

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#4 ylu

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Posted 09 September 2019 - 04:26 AM

Thank you for your suggestions.  I found a paper published on hematology. The s6k western also showed two bands.They didn't explain this.

 

 

 

 

hep26930-fig-0002-m.jpg







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