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Storage condition of Recombinant protein (native)


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5 replies to this topic

#1 Adrian K

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Posted 21 April 2011 - 08:05 AM

Dear All,

I would like to know your opinion about the best condition to store your recombinant protein.
I had recently expressed my recombinant proteins, sonicated and purified using his-tag column, and intend to do some functional studies later.
However I am puzzled on about what would be the best condition to store it.

Some says store in 4C , some say -20C, some says -80C. Some suggest to store with protease inhibitors.

Perhaps you may have some advice for me? I need your opinion.

Many thanks.

Adrian
Expecting the world to treat you fairly because you are a good person is like expecting the lion not to attack you because you are a vegetarian.

..."best of our knowledge, as far as we know this had never been reported before, though I can't possible read all the published journals on earth, but by perform thorough search in google, the keywords did not match any documents"...

"what doesn't kill you, makes you stronger"---Goddess Casandra reminds me to be strong

"It's all just DNA. Do it."---phage434

#2 mdfenko

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Posted 21 April 2011 - 11:02 AM

it depends on the protein. see if you can find the storage conditions for the native protein and use those.
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#3 Adrian K

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Posted 21 April 2011 - 06:52 PM

Is it safe to say that -20C will be the general "safe" condition? this is because (i heard) -80C and from water crystal which destroy the bonding of the protein, and freeze thaw will cause reduction of the protein quality...

or everything should just be a trial and error?

Thanks...
Expecting the world to treat you fairly because you are a good person is like expecting the lion not to attack you because you are a vegetarian.

..."best of our knowledge, as far as we know this had never been reported before, though I can't possible read all the published journals on earth, but by perform thorough search in google, the keywords did not match any documents"...

"what doesn't kill you, makes you stronger"---Goddess Casandra reminds me to be strong

"It's all just DNA. Do it."---phage434

#4 lab rat

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Posted 21 April 2011 - 07:05 PM

-20 should be OK as long as you leave it there until you need it. Aliquot the sample into single-use volumes and discard what you don't use. If it's a native protein, it should be fine in whatever elution buffer you used plus protease inhibitors. If you're not sure what your downstream application is, leave out EDTA.
42..."An immutable fixed-precision number of unlimited magnitude." <a href="http://en.wikipedia....amming_language)" target="_blank">http://en.wikipedia....amming_language)</a>, accessed 25June2009.

#5 protolder

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Posted 22 April 2011 - 06:28 AM

Hola, Adrian, firstly congratulations, I hope that you can write a lot of papers (I have read some of your blogs). Well, in my lab,people buy lots of tagged kinases for activity assays and all of them are dissolved in a buffer witth tris,7.5-8.0 50mM, NaCl 150mM a bit(0,1% Tx100) of detergent and reducer(1mM BME) and 50% glicerol and they are supplied and stored at -80ºC. I´m remembering it because I´m not in the lab, but check the storage conditions of any of the Kinases that Invitrogen sells. Thermo has a 4x stabilization solution for pure proteins. Buena suerte y mucho éxito profesional.

Edited by protolder, 22 April 2011 - 06:29 AM.


#6 Adrian K

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Posted 22 April 2011 - 10:25 AM

Hola, Adrian, firstly congratulations, I hope that you can write a lot of papers (I have read some of your blogs). Well, in my lab,people buy lots of tagged kinases for activity assays and all of them are dissolved in a buffer witth tris,7.5-8.0 50mM, NaCl 150mM a bit(0,1% Tx100) of detergent and reducer(1mM BME) and 50% glicerol and they are supplied and stored at -80ºC. I´m remembering it because I´m not in the lab, but check the storage conditions of any of the Kinases that Invitrogen sells. Thermo has a 4x stabilization solution for pure proteins. Buena suerte y mucho éxito profesional.



Mdfenko, LabRat and Protolder, thanks for all the input and sharing. :D
:lol:
Haha protolder, don't congratulate me: things was not turned out smooth lately.

The said paper had to change totally, so as the second one: the whole paper renovation. My bosses was not impressed, at all. Not only I have to re-write, but I have to do more work, wait for samples from overseas etc. But yet, those were not my thesis work, said my boss. For my thesis work, there are still more things to do, which I have not able to do yet. My research grant expired and frozen, I applied for the grant extension, but there is no news yet and the server had been down for a week. Geez, my stipend stopped starts from this month. I have to rely on my own to survive.

Everything just turned out to be the expected worst. :(

Edited by adrian kohsf, 22 April 2011 - 10:26 AM.

Expecting the world to treat you fairly because you are a good person is like expecting the lion not to attack you because you are a vegetarian.

..."best of our knowledge, as far as we know this had never been reported before, though I can't possible read all the published journals on earth, but by perform thorough search in google, the keywords did not match any documents"...

"what doesn't kill you, makes you stronger"---Goddess Casandra reminds me to be strong

"It's all just DNA. Do it."---phage434




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