- the protein is stable between pH 4.0 and 9.0
- FBPase is heat stable to 80 °C
- has a pI of 6.54
- has a relative molecular mass of 36770 Da.
So I've been asked this question, and I dont know why im so stumped. This is what i have so far:
To start, you would need e.coli to be lysed so the addition of would do this lysozyme (this works on gram negative bacteria in this scenario), following this bacterial proteases inhibitors would need to be added to stop proteases from breaking down the proteins as our protein of interest would go.
And this is where is all goes crap for me, like i know what needs to be done, but i do not know how to put it. I know that i need to run an SDS PAGE, i know that i can use western blotting but i have no idea what comes before what/
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Outline a purification strategy for FBPase from E. coli. watch
- Thread Starter
- 22-02-2018 20:36
- 22-02-2018 21:10
Look into chromatography.
Add an enzyme assay.
Throw in some mass spectrometry if you really want to go for overkill.