Horns like appearance on SDS-PAGE Silver stained Gel - (Jun/29/2010 )

Hey people... anyone ever observed such a thing on SDS-PAGE gel...
Its a non-reduced sample...
can u suggest reasons as to why this might happen??!!! is the salt concentration (1M NaCl) of the sample playing any role here??!!! i get these kind of profile in 140 mM salt also!!!

I run at 130V constant for one hour!!
Its a Lamellei buffer system!!

any furtehr details i can provide if u need!!!

Are you running these through a stacking gel? What % polyacrylamide are you using?

Looks overloaded to me.

leela on Jun 30 2010, 11:59 PM said:

4% stacking
4-15% gradient separating!!

DRT on Jul 1 2010, 05:41 AM said:

well its 10 micrograms!!! wud u cal it over loading!!!
cause for other samples on the same gel i dint get such a profile which were also 10 mcg!!!

Prep! on Jul 1 2010, 12:11 AM said:

if you are expecting a single band and are silver staining then yes.

DRT on Jun 30 2010, 07:11 PM said:

Agreed.

well since most of u feel its over loading i will do the next gel in varying amounts and get back to ya!!!
and md ya i do expect higher and lower molecular weight impurities and so the loading amount!!!