7 replies to this topic

[Prep!]

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!!!  

Attached Thumbnails

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Edited by Prep!, 29 June 2010 - 12:40 AM.

[leela]

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

[DRT]

Looks overloaded to me.

[Prep!]

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

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

4% stacking
4-15% gradient separating!!

[Prep!]

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

Looks overloaded to me.

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!!!

[mdfenko]

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

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

[HomeBrew]

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

Looks overloaded to me.

Agreed.

[Prep!]

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!!!