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Enzyme storage in eppendorphs to use in enzyme assay next day


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6 replies to this topic

#1 San1986

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Posted 05 August 2012 - 10:06 AM

Hi,

I have another question too. I am working on Cellulases from Bacteria. Is it possible that i centrifuge the culture medium in eppendorphs to get the crude enzyme but use this crude enzyme next day to perform assay for different parameters in its charaterization? If i keep this crude enzyme in refrigerator for a night, will it change the nature of the enzyme?

San.

#2 bob1

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Posted 05 August 2012 - 01:52 PM

Quite possibly it will change, it depends on how well inhibited the native proteases and other degrading enzymes are, and how stable the enzyme you are looking at is.

#3 San1986

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Posted 06 August 2012 - 11:52 AM

I have read in the literature to store the purified enzyme (post-ammonium sulphate precipitation procedure for proteins) at 4 C because the purified enzyme is unstable. So, if instead of the refrigerator, if i store the crude enzyme at 4 C for the next day use, will the possibilities of native proteases inhibiting the enzyme still be there?

I actually, want to save my time by acquiring crude enzyme by producing it on optimised conditions once for all, for the use in characterization and later on for purification by Ammonium sulphate precipitation method.

San

#4 bob1

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Posted 06 August 2012 - 04:43 PM

Purified enzymes should be fine at 4 for a while, but it depends on how pure they are and how stable they are.

#5 San1986

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Posted 07 August 2012 - 10:53 AM

Okay.
And how can i know if this is a stable enzyme or not? by performing assays?

#6 bob1

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Posted 07 August 2012 - 01:58 PM

The only way is to test them.

#7 San1986

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Posted 09 August 2012 - 08:06 AM

Okay thanks :)




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