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Protein detection


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3 replies to this topic

#1 uttess

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Posted 08 May 2011 - 07:28 PM

hello,

am looking for a protein of nearly 35 kDa via Western blot for which i use specific antibody...this time when i performed western, apart from this band (which was weak) i saw a dark band of about 60 kDa..the antibody is said to be specific...does anyone know how to account for this?

#2 mdfenko

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Posted 09 May 2011 - 08:12 AM

could be an artifact which shows up when using aged 2-mercaptoethanol (and dust). it will stain non-specifically.
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#3 Prep!

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Posted 09 May 2011 - 07:57 PM

hey md will the dtt artifact show up in western too? i agree abt the 60 kDa belonging to dtt artifact or dust and it does show up in my silver stained gels but they get eliminated during western!!!
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#4 uttess

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Posted 23 May 2011 - 02:28 PM

could be an artifact which shows up when using aged 2-mercaptoethanol (and dust). it will stain non-specifically.


thnx for the reply but i have bis tris gel and pvdf membrane, no silver..will it be an artifact?




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