RE:Cyclin E

I am using Cyclin E from NeoMarkers(Labvision) with a placental control.  I do
see the nuclear staining that is a true positive but I am also seeing a fairly
good amount of cytoplasmic staining as well.  I have titrated this antibody out
until there is no staining at all as well as running a negative control(which is
beautifully clean).  Has anyone else seen this and if you have how have you
treated this?  I am using LSAB+ kit from Dako with DAB.

I'm also looking for ways to get rid of background on human kidney.  I have
avidin/biotin blocked for 15 minutes each but the negative control still has
very specific background.  Should I switch to an alk phos system?

Christine Klein
MD Anderson Cancer Center.

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