Melanie

Hi, you are not alone dear! These 2 are the most temperamental antibodies
one could come across. They have minds of their own, I guess it also
depends on the tissue type and fixation protocols.
I have used Novocastra's {NCL-4C7-CD5 }: MW pretreatment for 20 mins in 1mM
EDTA pH 8.0 - ab dilution 1:25 1 Hr at rt' and the CSA kit ( DAKO) for
detection
and
{NCL-CyclinD1-GM} clones; MW in Tris base pH 10.0  for 10 mins or 20 mins
whichever works for you. Ab dilution 1:50 overnight at rt' and detection
using DAKO CSA kit.
Try it ; go for gold! It may just work for you.
Good luck.

Cheers

Zenobia Haffajee
HAPS, Newcastle, 2300.