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protein optimization


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3 replies to this topic

#1 kentz19

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Posted 05 June 2010 - 06:43 AM

Hi everyone,

I'm working on enzyme assay. My question is, do I need to optimize the enzyme assay condition again once I used a new batch of protein(microsomes). The procedure of preparation is the same as before and the protein content is almost the same. Do I need to do optimization again or just proceed with the experiment using optimization data from previous batch of microsomes?

#2 Inmost sun

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Posted 06 June 2010 - 02:48 AM

Hi everyone,

I'm working on enzyme assay. My question is, do I need to optimize the enzyme assay condition again once I used a new batch of protein(microsomes). The procedure of preparation is the same as before and the protein content is almost the same. Do I need to do optimization again or just proceed with the experiment using optimization data from previous batch of microsomes?



if you are satisfied with your current protocol you should use it routinely; if not start another optimization

#3 oasis

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Posted 06 June 2010 - 11:53 PM

hi,there
it would be more useful and accurate to optimise each lot to avoid any sort of problem.

#4 NHI

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Posted 13 July 2010 - 12:20 AM

hi fren...

this is my suggestion, there is a few condition and situation that u need to remember...

1) Locked-Process sample generation: means, u lock all ur process from upstream and and downstream including ur analytical methods once u already have a consistent parameter

2) Development sample: if ur sample is not consistent, i suggest you to test both. i.e. first u need to use a locked-method for analysis and another secondly, u need to develop the optimize analytical method if u think its really needed...

by the way, normally people lock the analytical method (after doing their semi optimise method) throughout the development....in later stage, they will double confirm the method by using the consistent batch sample

this is juz from my own experience..tq




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