I performed a native gel electrophoresis after an extraction in native lysis buffer on C.elegans. But something is "stuck" in the gel (where the 26s 1 cap should be seen).I have used a 4% gel.
thanks !
It would be nice if you could give some more details on the protocol you used.
For example:
- Did you perform BN-PAGE (= adding Coomassie Blue in the sample buffer) or just regular PAGE?
- Composition of the native lysis buffer?
- Running conditions?
- Did you cast your own gel or did you buy one?
Posting a picture of the gel could also be useful.
Ok sure. So, I added in the samples 2µL of blue ( glycérol + electrophoresis buffer + xylene cyanol).
The composition of lysis buffer : 20mM Tris Hcl, 5mM MgCl2, 0.5% NP-40, 10% glycerol, 1mM ATP, 1mM DTT. pH 8
I ran the gel in a cold room for 5h at 100V
And I casted the gel (3mL Tris-borat 5X, 1.5mL acrylamide 40%, 15µL DTT 1M, 150%L ATP 100mM, 120µL APS 10%, 12µL TEMED, qsp 15mL of water)
See here a picture of the gel
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