Hello,
I'm trying to figure out how long is the DTT good for. I know some people store the loading sample buffer containing DTT in the -20 and use it. Will this be good, or is it better to add DTT every time u use loading buffer (for western blot). And if I have to add it new every time what is the best way to do this. Weighing the grams of DTT and adding it to the loading buffer, or can I have a solution stock?
Right now I have 2x loading buffer with DTT that I stored in -20C I want to use it but I don't know if I have to add fresh DTT. I also boil the sample 5min @ 95C.
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#2
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Posted 31 August 2009 - 08:50 AM
medchemgirl, on Aug 31 2009, 11:31 AM, said:
Hello,
I'm trying to figure out how long is the DTT good for. I know some people store the loading sample buffer containing DTT in the -20 and use it. Will this be good, or is it better to add DTT every time u use loading buffer (for western blot). And if I have to add it new every time what is the best way to do this. Weighing the grams of DTT and adding it to the loading buffer, or can I have a solution stock?
Right now I have 2x loading buffer with DTT that I stored in -20C I want to use it but I don't know if I have to add fresh DTT. I also boil the sample 5min @ 95C.
I'm trying to figure out how long is the DTT good for. I know some people store the loading sample buffer containing DTT in the -20 and use it. Will this be good, or is it better to add DTT every time u use loading buffer (for western blot). And if I have to add it new every time what is the best way to do this. Weighing the grams of DTT and adding it to the loading buffer, or can I have a solution stock?
Right now I have 2x loading buffer with DTT that I stored in -20C I want to use it but I don't know if I have to add fresh DTT. I also boil the sample 5min @ 95C.
I keep aliquots of DTT stored at -20°C, and add freshly thawed DTT to my loading buffer every time with good success.
Science is simply common sense at its best that is rigidly accurate in observation and merciless to fallacy in logic.
Thomas Henry Huxley
#3
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Posted 31 August 2009 - 12:53 PM
Thanks!
I read that DTT gets oxidized very quickly even when stored at -20, so I was confused about if I could still use it. Do you have 1M stock or is it more concentrated?
I keep aliquots of DTT stored at -20°C, and add freshly thawed DTT to my loading buffer every time with good success.
I read that DTT gets oxidized very quickly even when stored at -20, so I was confused about if I could still use it. Do you have 1M stock or is it more concentrated?
Dr Teeth, on Aug 31 2009, 10:50 AM, said:
medchemgirl, on Aug 31 2009, 11:31 AM, said:
Hello,
I'm trying to figure out how long is the DTT good for. I know some people store the loading sample buffer containing DTT in the -20 and use it. Will this be good, or is it better to add DTT every time u use loading buffer (for western blot). And if I have to add it new every time what is the best way to do this. Weighing the grams of DTT and adding it to the loading buffer, or can I have a solution stock?
Right now I have 2x loading buffer with DTT that I stored in -20C I want to use it but I don't know if I have to add fresh DTT. I also boil the sample 5min @ 95C.
I'm trying to figure out how long is the DTT good for. I know some people store the loading sample buffer containing DTT in the -20 and use it. Will this be good, or is it better to add DTT every time u use loading buffer (for western blot). And if I have to add it new every time what is the best way to do this. Weighing the grams of DTT and adding it to the loading buffer, or can I have a solution stock?
Right now I have 2x loading buffer with DTT that I stored in -20C I want to use it but I don't know if I have to add fresh DTT. I also boil the sample 5min @ 95C.
I keep aliquots of DTT stored at -20°C, and add freshly thawed DTT to my loading buffer every time with good success.
#4
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Hmmm, I think it's working
Posted 31 August 2009 - 02:12 PM
We use DTT containing loading buffer straight from the -20, it works fine. Some of out loading buffer has been stored for a year or more.
#5
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Posted 01 September 2009 - 05:26 AM
bob1, on Aug 31 2009, 06:12 PM, said:
We use DTT containing loading buffer straight from the -20, it works fine. Some of out loading buffer has been stored for a year or more.
Good to know. In response to an earlier question, we store our DTT as 1 M stock.
Science is simply common sense at its best that is rigidly accurate in observation and merciless to fallacy in logic.
Thomas Henry Huxley
#6
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Posted 01 September 2009 - 06:39 AM
DTT has an absorbance peak of 220 nm, while the oxidized form has a peak at 300 nm.
