Stability of IPTG, DTT at 4 degree - (Nov/19/2008 )
hello fellow researchers,
i am using IPTG stock made 50 ml at a time. I want to know whether repeated thawing and freezing affects IPTG stability.
Also, before induction if i keep the IPTG at 4 degree for 4-5 hrs does this affect is stability. I am facing problem with induction in BL21 strain.
One more thing is I am using NuPAGE sample buffer . This buffer needs DTT. I have prepared and stored this buffer at 4 degrees. Will this have affect on the NuPAGe sample buffers ability to denature proteins.
-abhijimmy-
QUOTE (abhijimmy @ Nov 19 2008, 06:10 AM)
hello fellow researchers,
i am using IPTG stock made 50 ml at a time. I want to know whether repeated thawing and freezing affects IPTG stability.
Also, before induction if i keep the IPTG at 4 degree for 4-5 hrs does this affect is stability. I am facing problem with induction in BL21 strain.
One more thing is I am using NuPAGE sample buffer . This buffer needs DTT. I have prepared and stored this buffer at 4 degrees. Will this have affect on the NuPAGe sample buffers ability to denature proteins.
i am using IPTG stock made 50 ml at a time. I want to know whether repeated thawing and freezing affects IPTG stability.
Also, before induction if i keep the IPTG at 4 degree for 4-5 hrs does this affect is stability. I am facing problem with induction in BL21 strain.
One more thing is I am using NuPAGE sample buffer . This buffer needs DTT. I have prepared and stored this buffer at 4 degrees. Will this have affect on the NuPAGe sample buffers ability to denature proteins.
Hi, IPTG at 4-5 hours should not affect the stability much. Also, storing DTT at 4 degrees for this time will not either. Is there a reason that you cannot freeze aliquots of your chemicals? This is much better to do because both of these chemicals will reduce in solution after a period of time and become ineffective.
What kind of problems are you having with induction? Have you run an induction test? Do you see little induction, or none at all? This could be the root of many problems not having anything to do with IPTG.
Let us know! Good luck!
Cindy
-biokmst-