I have been trying to analyze some embryonic zebrafish
proteins using 1D-SDS-PAGE and western blotting, but have
experienced a very upsetting problem with some of them. I am
currently detecting bands (on the blot) as if they have
never entered the separation gel matrix. I have worked with
orthologs of this protein and the predicted sequences show
high identity so I would expect them to migrate properly.
Anyone has gone through a similar trouble? Do you think this
could be due to protein extraction and contamination with
extracellular matrix components? BTW, I'm extracting with
Triton X100 so I wouldn't expect significant DNA
contamination, nor yolk as I am manually deyolking before
homogenizing.
Thanks for your help,
D
_______________________________________________
Methods mailing list
[email protected]
http://www.bio.net/biomail/listinfo/methods
