w******y
发帖数: 2504 | 1 The PVDF WB membrane was first probed with several different antibodies and
stripped between each probing, then it was stored in TBST. Yesterday, I
tried to probe for another target protein, seems like this old membrane dose
not work any more even if I probed with the antibody that worked on this
membrane. Any suggestions for this situation? Thanks. | s******y
发帖数: 28562 | 2 I suggest you don't bother.
It is likely all the protein on the surface have been degraded due to
bacterial activity and other factors.
Well, however, if you are really, really,really desperate, you can try this:
wet the membrane in methanol for 5 minutes, then wash by PBS three times.
Then maybe you can try again.
Remember: if you want to keep a membrane for a long time, always dry it, and
then store at -20oC
and
dose
【在 w******y 的大作中提到】
: The PVDF WB membrane was first probed with several different antibodies and
: stripped between each probing, then it was stored in TBST. Yesterday, I
: tried to probe for another target protein, seems like this old membrane dose
: not work any more even if I probed with the antibody that worked on this
: membrane. Any suggestions for this situation? Thanks.
| w******y
发帖数: 2504 | 3 Thanks so much, Sunnyday. | s******y
发帖数: 28562 | 4 Also, if you have to do this, you need to use the most sensitive detection
reagent,
such as the Western Femto detection kit from Pierce Chemical
【在 w******y 的大作中提到】
: Thanks so much, Sunnyday.
|
|
