SDS PAGE

[Clint P]

Hey everyone-

 

This is basically just a curiosity question. We ran a column and an SDS page for protein separation. We separated Ferratin, BSA, and Cyt C (they eluted in that order). Here is a pic of my annotated SDS page:

 

 

 

 

(the last 4 wells on the right are unknowns, so that is why they are out of order)

 

Also, the scale is arbitrary, 0=start, 100=finish

 

Does anyone know what the wells marked (21) could be? They appear to be 180ish kDa according to the ladder. However, ferratin subunits are 20kda, BSA is 66kda, and cyt c is 12kda. Any ideas? They appeared to have eluted at the same time as BSA.

 

Thanks!

[ewmon]

I'm no SDS PAGE expert, but the 33 bands looks more like the 88 kDa species of BSA, and the 21 band is more like ~190 kDa but I have no idea what it could be.

[CharonY]

I assume you mean ferritin? What kind of column was used (not pertaining to the question really, but it is important to realize that you have a gazillion of column materials and running a column carries much less information than, e.g. saying one is running an SDS gel). That being said, depending on the preparation method albumin tends to multimerize. These tend to be very stable and are not easily resolved even in a denaturing gel.

 

With better resolution you are likely to see also bands at e.g. 130 and around 190. Tip: use marker left and right of your samples to better assess smiling effects.

Edited March 23, 2012 by CharonY