Dear Jacob,
somewhat adding to list of 'not really answering your question', here is
the reference for native page that still uses a dye thereby trying to
limit the influence of the charge on the speed. Might be helpful as they
discuss some applications. Otherwise AUC really sounds like the way to go.
Nat Protoc. 2006;1(1):418-28.
Blue native PAGE.
Wittig I, Braun HP, Schägger H.
best,
hannes uchtenhagen
Hannes Uchtenhagen
Karolinska Institutet
Center for Infectious Medicine (CIM)
Karolinska University Hospital Huddinge, F59
SE-141 86 Stockholm, Sweden
Office: +46-(0)8-524 86981
Mobile: +46-(0)7-36901461
On 2010-05-18 22:04, Jacob Keller wrote:
Dear Crystallographers,
I am trying to optimize a native gel experiment of a two-protein
complex, running the smallest-detectable amount of protein component A
with varying amounts of component B.
MW Charge MW/Charge
A 22 -5 -4308
B 17 -24 -702
This experiment is partly to determine stoichiometry, but also to
determine roughly the strength of the interaction.
B definitely runs much faster than A alone, as predicted, but I am
wondering what to expect with various oligomers. Should ABB run faster
or slower than AB? What about AABB? Theoretically, AA should certainly
run slower than A, and BB slower than B, simply because the mass/charge
ratio is the same, but the overall mass is greater. But what happens
when you have AAB, for example? There must be an equation relating the
mass/charge and mass (and perhaps gel percentage) to the speed traveled
in the gel--but what is the equation?
Thanks for your consideration,
Jacob
*******************************************
Jacob Pearson Keller
Northwestern University
Medical Scientist Training Program
Dallos Laboratory
F. Searle 1-240
2240 Campus Drive
Evanston IL 60208
lab: 847.491.2438
cel: 773.608.9185
email: j-kell...@northwestern.edu
*******************************************
