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I'm analysing cultured fibroblasts by western blotting (protein extracted in RIPA buffer) and complex I/IV dipstick assays (in buffer A from ab109876 or ab109720). I'm wondering if it's possible to use the buffer A samples for western blotting? if yes, any additional steps required before loading or it's the same like how i treat my RIPA samples?
Asked on May 21 2012
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Yes these samples can be used for Western blotting just as one would with RIPA, the extraction detergent in A is very similar to RIPA (NP40 plus others) but it is more native.
The only thing that I would say is that when used concentrated and diluted in the Western blot sample buffer, detergent may affect SDSPAGE migration at the very bottom of the lane (much less than 10kDa) but all other migration should be normal.
I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.
Answered on May 21 2012