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Inaccuracies in protein MW on SDS-PAGE
Hello everyone, got a quick question about the reliability of MW estimations using SDS-PAGE. I've got a protein which is estimated to be ~60 kDa, although on my gel it seems to consistently migrate closer to 65 kDa.
I know that SDS-PAGE may not always be accurate for measuring these things, but is there any way to improve on this? What is the reason for the higher migration? Improper denaturation?
Re: Inaccuracies in protein MW on SDS-PAGE
There are a lot of possible explanations.
First of all are you using a pre-stained marker?. Pre-stained marker gives an approximate idea of the molecular but unstained markers are more precise. May be you could run BSA (Molecular weight: 66382) in one well to check if your marker is behaving good.
Other options are:
-your protein may have post-traductional modifications (glicosilation, phosphorilation, an so on)
-your loading buffer could be interfering with your run.
|inaccuracies , protein , sdspage|
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