Cyclin D1

From:"Brett, Lawrence"

Rita
This is quite a difficult antigen and requires intense antigen retrieval. We've found a combination of Pressure cooking in Tris-EDTA pH8.0 followed by 15 seconds in 0.05% Pronase E to give good results with the Novocastra antibody at 1:50. You can also use a high pH AR solution like Dako's 'High pH Target Rerieval Solution' which is pH 9.9.
Detection is PowerVision which is more sensitive than most other systems.
Hope this helps
 
Lawrence Brett
Royal Infirmary, Edinburgh. UK
 
 
_____________________________________________________________
Dear Histonetters,
 
I am  trying to get an optimum staining technique for Cyclin D1 on bone
marrow trephines.
Any advice as to what you are doing to make this difficult stain
reproducibly and reliably accurate? (antibodies, retrieval, fixation, kits,
timing, etc..) Any good ideas or hints would be much appreciated.
Many thanks from Down Under!
 
Rita Broner
Bone Marrow Transplant Unit
Alfred Hospital
Melbourne, Australia 


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