Does anyone have a protocol on how to elute proteins from SDS-PAGE gel bands?
Thanks!
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oana
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Posted 08 July 2002 - 07:54 AM
Hi Engen!
I was interested in the same protocol and I manage with this one:
-make a preparative gel and cut off the band which contain your protein of interest (I visualised the gel in 0.1M KCl, 30 min,4C)
-try to slice the band you cut off in as small pices as you can (I frozen the paa and then I sliced it)
-add elution buffer (1ml to 400mg paa) and let the protein to be eluted over night at RT, on a shaker
elution buffer: 20mM TrisHCl pH 7.7, 150mM NaCl, 0.1% SDS, 2mM EDTA
-then you can check the elution in SDS-PAGE
I was interested in the same protocol and I manage with this one:
-make a preparative gel and cut off the band which contain your protein of interest (I visualised the gel in 0.1M KCl, 30 min,4C)
-try to slice the band you cut off in as small pices as you can (I frozen the paa and then I sliced it)
-add elution buffer (1ml to 400mg paa) and let the protein to be eluted over night at RT, on a shaker
elution buffer: 20mM TrisHCl pH 7.7, 150mM NaCl, 0.1% SDS, 2mM EDTA
-then you can check the elution in SDS-PAGE
For me it worked. Good luck!
Oana
