Solubilization of Triton insoluble pellet - (Dec/23/2008 )
Hi,
Does anyone has experience in solubilizing Triton X 100 Insoluble pellets? I would really appreciate if I could get your inputs.
I tried boiling the pellet in 1X/ 2X sample buffer and yet nothing happened.
Thanks,
Tulip
-tulip-
have you tried urea?
the pellet, if it is protein, should dissolve in sds-page loading buffer. maybe you tried to dissolve too much for the volume?
also, try lower than boiling temperature (65C should be okay) to avoid aggregation (it will take longer to dissolve).
-mdfenko-
QUOTE (mdfenko @ Dec 24 2008, 11:38 AM)
have you tried urea?
the pellet, if it is protein, should dissolve in sds-page loading buffer. maybe you tried to dissolve too much for the volume?
also, try lower than boiling temperature (65C should be okay) to avoid aggregation (it will take longer to dissolve).
the pellet, if it is protein, should dissolve in sds-page loading buffer. maybe you tried to dissolve too much for the volume?
also, try lower than boiling temperature (65C should be okay) to avoid aggregation (it will take longer to dissolve).
Thank you for your suggestion. What concentration of Urea should I be using to dissolve the pellets?
Tulip
-tulip-
6-10M. we usually use 8M.
-mdfenko-