Posted 03 September 2010 - 04:03 AM
Posted 03 September 2010 - 05:29 AM
Check for truncation of your recombinant protein, As if you have N terminal tag u will get both truncated and untrancated protein. SO please check your sequencing results for a mutation which leads to Truncation or Try to grow cells at lower temperature and Induce with lower IPTG or other inducer.
Posted 03 September 2010 - 08:20 PM
genius does what it must
i do what i get paid to do
Posted 11 September 2010 - 08:48 AM
Posted 14 September 2010 - 07:27 AM
Another possibility is phosphorylation which may cause an upshift in your gel. Bacteria contain machineries that may phosphorylate your protein from human or whatever species. You can dephosphrylate you sample and see if the band shift back down.
Posted 21 September 2010 - 10:09 PM
why dont u try a western... may be the 30 kDa protein is a major contaminant if its not your product!!
Edited by Prep!, 21 September 2010 - 10:11 PM.
Posted 22 September 2010 - 07:49 PM
Posted 22 September 2010 - 07:52 PM
as pointed out earlier... SDS-PAGE is a best estimate of the molecular weight but not necessarily the accurate one!!
reason of editing: spell-check!!
Edited by Prep!, 22 September 2010 - 07:52 PM.