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Enzyme assay


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#1 mahroo1356

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Posted 02 January 2013 - 07:56 AM

Hi and happy new year,

I am going to prepare pyruvte kinase enzyme for section of assay process of starch synthase enzymic activity .
Pyruvate kinase enzyme is as a crystalline suspension in 3.2 M ammonium sulfate, pH 6.0. (5000 unit ).
for each assay 1.2 unit of this enzyme must be used. I know how calculate the amount of µL that is need for each assey and i know how prepare the work stock, but i don't know how can i prepare an solution of purchased enzyme as purified stock. I want to know the preperation method of purified stock of enzyme that is as a crystalline suspension. (form of purchased enzyme is crystalline particles). Prompet reply will be really appreciated.

#2 bob1

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Posted 02 January 2013 - 02:10 PM

Check the product sheet (often available online if it is a commercial company) for the correct conditions for re-suspending the enzyme. The resuspension will probably be in a buffer solution of some sort and the sheet should tell you storage conditions.

#3 mahroo1356

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Posted 02 January 2013 - 10:08 PM

Check the product sheet (often available online if it is a commercial company) for the correct conditions for re-suspending the enzyme. The resuspension will probably be in a buffer solution of some sort and the sheet should tell you storage conditions.


Dear bob 1, first of all thanks for your prompt reply, I attached data sheet, but i couldn't find correct method for resuspension in that. I solved 1mg of this enzyme in hepse buffer (this bufer is incobation buffer in my test) but precipitation took place and some white powder remained on microtube. I really confused, would you please help me?

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#4 bob1

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Posted 03 January 2013 - 02:18 PM

Check out the references, especially number 4.




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