Peptide transfer to PDVF

ming at msvax.mssm.edu ming at msvax.mssm.edu
Tue Apr 23 09:24:55 EST 1996


First, you have to use a PVDF membrane which has a smaller pore size,
e.g., 0.2 or less. Second, time your transfer carefully, or put an additional
layer of membrane on the back of the first one. Third, if your page was not
a SDS style, you have to make a sandwitch to do the transfer and check
the both membranes simultaniously. At last, if you are only detecting
your peptides by staining, remember that almost all dyes have a less
sensitivity on PVDF than on the gel itself. Good luck!




In article <4l7iva$n3v at studium.student.umu.se>, Malgorzata Wilczynska
<margaret at solaris.chem.umu.se.> writes:
>
>
>Hey everybody,
>
>I have a mixture of peptide fragments from 2 kDa to 20 kDa. 
>I can see all these peptides on polyacrylamide gel, but after 
>transfer to PDVF membrane (Western blotting according to Towbin) 
>I can detect only fragments bigger than 9 kDa. Is it possible 
>that peptides smaller than 9 kDa do not bind to PDVF membrane? 
>If yes, does anyone know how to improve the transfer of peptide?
>                                    
>                                    Thanks in advance,
>                                    
>                                    Malgorzata Wilczynska
>                                    Medical Biochemistry
>                                    Umeå University
>                                    margaret at solaris.chem.umu.se.
>



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