SDS-PAGE running strangely
Posted 26 March 2005 - 07:28 AM
FEBS Lett. 1984 May 7;170(1):81-4.
SDS-PAGE strongly overestimates the molecular masses of the neurofilament proteins.
I also have situation similar to you. My protein is about 12kDa, but it appears as about 16kDa in SDS PAGE. My protein has a flexible and very negatively charged C-terminal tail, so i think the tail may hinder the mobility. The negative charge residues may also affect SDS binding to protein, thus lower the mobility.
Posted 05 March 2009 - 10:38 PM
Posted 12 March 2009 - 06:58 PM
is your gel heating up? try running at constant mAmps or lower the voltage
Posted 15 May 2009 - 08:14 PM
there might be possibility that the height of stacking gel is playing some role here. If the height is less than 1 cm than it can result poor resolution of protein markers.
Posted 03 August 2009 - 02:26 AM
ChiMy, on Mar 10 2005, 12:54 AM, said:
Is that the protein surface charge that involve in the migration of protein on SDS PAGE?
Thank you in advance.
Check your protein may be glycated. In some cases glycated protein show such strange behaviour
Posted 04 July 2012 - 07:38 AM
Posted 15 August 2012 - 07:10 AM
If you are using 6% gel, you won't be able to see those 10-20KD markers, or even 30KD markers if your gel is not that long.
You should choose wisely which concentration to use according to the molecular weight of the target protein.
The marker cannot run faster than the loading dye. But the chlorophyll can.
Posted 20 August 2012 - 05:23 PM
Sometimes proteins dont migrate just based on their MW - protein characteristics affect the migration