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Horns like appearance on SDS-PAGE Silver stained Gel


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7 replies to this topic

#1 Prep!

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Posted 29 June 2010 - 12:39 AM

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!!! ;)

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

Support bacteria - They are the only culture some people have!!!
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#2 leela

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Posted 30 June 2010 - 09:29 AM

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

#3 DRT

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Posted 30 June 2010 - 03:11 PM

Looks overloaded to me.

#4 Prep!

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Posted 30 June 2010 - 08:10 PM

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


4% stacking
4-15% gradient separating!!
Support bacteria - They are the only culture some people have!!!
Cheers!!!

#5 Prep!

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Posted 30 June 2010 - 08:11 PM

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!!!
Support bacteria - They are the only culture some people have!!!
Cheers!!!

#6 mdfenko

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Posted 01 July 2010 - 06:29 AM

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.
talent does what it can
genius does what it must
i do what i get paid to do

#7 HomeBrew

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Posted 01 July 2010 - 06:34 AM

Looks overloaded to me.


Agreed.

#8 Prep!

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Posted 01 July 2010 - 11:50 PM

well since most of u feel its over loading i will do the next gel in varying amounts and get back to ya!!! :lol:
and md ya i do expect higher and lower molecular weight impurities and so the loading amount!!!
Support bacteria - They are the only culture some people have!!!
Cheers!!!




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