I am using the recipe of mild strip buffer suggested by abcam.
http://www.abcam.com/ps/pdf/protocols/Stripping%20for%20reprobing.pdf
It works fine but after one month, the solution turned milky, I wonder whether it is normal and can the buffer still be used?
Hi Kc,
since your buffer contains SDS it might simply as gotten to cold leading to precipitation of SDS.
Try heating it shortly in a microwave or waterbath. If the turbidity vanishes it must have been the SDS. If not you should probably prepare a new buffer.
This buffer should be stored at room temperature, because of the SDS.
Best,
Kai
Hi Kc,
It could be the SDS precipitating out as Kai suggests, but it is only at low concentration (0.1% SDS) so this should not happen at room temperature.
The recipe I use is slightly different:
0.2 M glycine pH 2.5
1% SDS
I store this at room temp and have never known it go cloudy. I incubate the membrane at room temp for 10-15 min with gentle shaking then wash with TBST and start with re-blocking. For re-probing I add 0.02% sodium azide to the blocking step to kill off any residual HRP activity from the previous ECL step (assuming you're detecting by ECL).
Hope that helps,
Mark
We have had the same problem, and heating the buffer (200 mL) for about 30 seconds in the microwave solved it. So it seems that our air conditioning was too low, that it precipitated the SDS.
The low amount of SDS isn't the main/only reason that the solution turned cloudy. It does contribute, however, note that the solution is at a very low pH. This causes the salt to precipitate, hence the milky solution. Once it turns milky, it can't be used for effective stripping (i've tried that before and it doesn't strip effectively). Instead, prepare a new one and instead of preparing at 1 L, prepare a smaller volume instead (:
- Badges
- Science method