Question (63117) | EPO overexpression 293T lysate (whole cell) (ab94196)
Go to datasheet (ab94196)We have done a few western blots with this EPO expressing cell lysate and don't get the expected band at around 34kDa; just see a band around 40 and 60kDa;
1. Do I have to add SDS-PAGE loading buffer to protein extract prior to loading on gel or is this a ready to load control?
2. Is it recommended to heat this sample prior to loading? If so, what is recommendation?
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Asked on Oct 08 2012
This lysate is ready to use and can be loaded directly to the gel.
Typically, 15uL of the lysate is loaded per lane for western blotting assays.
Our R and D department suggests following these guidelines. For some proteins, it may be necessary to heat the lysate at 95 degrees C for 3 minutes prior to loading the gel.
I look forward to hearing if this information proves useful. Please feel free to contact me again with any further questions.
Answered on Oct 08 2012
