PVDF direct blot/sequencing

Bill wnowatzk at ucs.indiana.edu
Tue Jan 25 14:01:27 EST 1994


I have had a bit of trouble transferring my 95 kD protein from a 10%SDS
PAGE to a BioRad PVDF membrane. I am considering trying a direct blot
method mentioned in the BioRad PVDF mb info booklet and further referenced
to David Speicher. I am wondering if anyone has experience with this
technique and what kind of success you have achieved upon sequencing? I
could also use some experimental condition recommendations- should I dilute
my protein (currently in a Tris buffer at about 2.4 mg/mL) into 200 µL and
rotate this in a small eppendorf tube containing a small strip of PVDF
overnight at 4 C? I am also curious to know if certain buffers interefere
with N terminal sequencing? Thanks in advance.

wnowatzk at ucs.indiana.edu



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