Dear All:
I'm testing a mdm2 anti-human antibody (OP115 Calbiochem, 2A10 clone) in FFPE human samples. I Have a suitable protocol for breast carcinoma, skin and colon but when I try it in liver metastasis I have a lot of background in the liver, I have another protocol that cleans this background but then the staining in colon/metastasis is not so intense and specific in the nucleus/cytoplasm.
Does anyone have experience with this antibody? I have to stain a TMA so I need a protocol suitable for all tissues.
Protocol 1; (skin, colon and breast carcinoma): Blocking steps: 10 minutes of TBST (Triton 0,1%),10 minutes 3% H2O2 n PBS, 30 min of 5% normal goat serum in PBS. HIER: MW 20 minutes at 600W (10 minutes boiling), cool 20 min a RT. Primary antibody overnight at 4ēC 1:150, 30 minutes of ENVISION anti-mouse, vector VIP substrate (purple).
Protocol 2(liver): Same as protocol 1 but using DAKO retrieval solution Ph:6
I can not attatch images...sorry, I know that this question is for the IHC forum but imposible to post there too...
Thanks in advance
Best regards
