inclusion bodies - (Nov/27/2007 )
Hi,
I am currently working on the purification of recombinant proteins of mycobacteria, i hav some doubts regarding this.Hope somebody will help me.
Will imidazole,NaCl,urea etc in the elution buffer interfere with elisa performance of these proteins?
If so how to remove these? i read abt dialysis, but i also read removal of NaCl, causes precipitation of protein in dialysis bags?
After elution of proteins, should we lyophilize these proteins, kindly tell me abt the procedures to be adopted after elution.
If we dont know the exact function of the genes, is there any software,online, to predict that?
Many thanks
-niveda-
QUOTE (niveda @ Nov 28 2007, 04:02 AM)
Hi,
I am currently working on the purification of recombinant proteins of mycobacteria, i hav some doubts regarding this.Hope somebody will help me.
Will imidazole,NaCl,urea etc in the elution buffer interfere with elisa performance of these proteins?
If so how to remove these? i read abt dialysis, but i also read removal of NaCl, causes precipitation of protein in dialysis bags?
After elution of proteins, should we lyophilize these proteins, kindly tell me abt the procedures to be adopted after elution.
If we dont know the exact function of the genes, is there any software,online, to predict that?
Many thanks
I am currently working on the purification of recombinant proteins of mycobacteria, i hav some doubts regarding this.Hope somebody will help me.
Will imidazole,NaCl,urea etc in the elution buffer interfere with elisa performance of these proteins?
If so how to remove these? i read abt dialysis, but i also read removal of NaCl, causes precipitation of protein in dialysis bags?
After elution of proteins, should we lyophilize these proteins, kindly tell me abt the procedures to be adopted after elution.
If we dont know the exact function of the genes, is there any software,online, to predict that?
Many thanks
Slightly confused here. Your title suggests you are working with inclusion bodies that you are resolubilizing, yes? If so, are you also refolding the proteins?
Depending on what you are doing, the short answer to your question could be 'Yes' or 'No' (sorry). Imidazole probably won't have any effect, but high levels of NaCl, urea etc most probably will. You'll need to desalt (GE column for example) or refold and reconcentrate.
-swanny-