Native PAGE, interaction of proteins, validity - (Jan/27/2012 )

Dear Protocol-Online

I am running a series of native PAGE experiments, using Bio-RAD TGX 4-15% gels, and Tris-Glycine buffer pH=8.1

My protein of interest has a pI of about 6, and I am investigating it's interactions with some multimeric proteins.

When running the experiment I usually prepare lanes as follows:
1)multimer alone
2)multime + protein
3)protein

I use silverstain for visualization.

I do not observe a direct colocalizaion of my protein on the multimers, but it surely changes its
pattern, and the protein in lane 2) does not wonder as far as the protein alone in 3).

What can I conclude from an experiment like this?

How du you like my setup for native PAGE in general?

And yeah, this is my first time with native PAGE.

best regards
Peter Jallerup

a picture would have helped but...

if lane 2 is slower moving than and there is no band at the same place as lanes 1 and 3 then you are seeing interaction.

your experimental design appears to be okay

Thank you very much for the answer

I know a picture would have been awesome, but my supervisor have read 1984 too many times.

best regards
PJ

PeterJallerup on Wed Feb 1 11:55:05 2012 said:

Sorry, what you mean by "read 1984"? I'm not a native English speaker...

Adrian K on Wed Feb 1 15:35:07 2012 said:

1984 is a novel, written by "george orwell" (pen name for eric arthur blair), an english author, about a future totalitarian state. he revealed the concept of the "big brother" government (among others). he also wrote "animal farm".