Help-IHC nuclear staining - (Aug/19/2010 )
I am dealing with IHC frozen section. Imet some difficulty of this antibody.My antibody is monoclonal and located at nuclear. It seemed it work fine for human colon cancer. But when I applied it on human ovarian cancer tissue.
When I stained the antibody, I found out that most of the malignant cells did not shown nuclear staining. There is some nuclear staining for malignant cells at the peripheral of tissue. I also noticed that this antibody also showed cytoplasm staning.
Is it possible to have this kind of staining? Or do there have any suggestion to improve better nuclear staining of the antibody for IHC frozen section?
You may need to do an antigen retrieval step. I have always done this on parrafin sections (not frozen) as the actual process (that I used) involves heat.
Clare
yellow18 on Fri Aug 20 04:06:19 2010 said:
I am dealing with IHC frozen section. Imet some difficulty of this antibody.My antibody is monoclonal and located at nuclear. It seemed it work fine for human colon cancer. But when I applied it on human ovarian cancer tissue.
When I stained the antibody, I found out that most of the malignant cells did not shown nuclear staining. There is some nuclear staining for malignant cells at the peripheral of tissue. I also noticed that this antibody also showed cytoplasm staning.
Is it possible to have this kind of staining? Or do there have any suggestion to improve better nuclear staining of the antibody for IHC frozen section?
Actually, I perfomed this antibody with FFPE sample (with antigen retrieval step of course). The result is more better but it still didn't showed satisfied nuclear staining 
Yellow18
Clare on Fri Aug 20 10:00:04 2010 said:
You may need to do an antigen retrieval step. I have always done this on parrafin sections (not frozen) as the actual process (that I used) involves heat.
Clare
yellow18 on Fri Aug 20 04:06:19 2010 said:
I am dealing with IHC frozen section. Imet some difficulty of this antibody.My antibody is monoclonal and located at nuclear. It seemed it work fine for human colon cancer. But when I applied it on human ovarian cancer tissue.
When I stained the antibody, I found out that most of the malignant cells did not shown nuclear staining. There is some nuclear staining for malignant cells at the peripheral of tissue. I also noticed that this antibody also showed cytoplasm staning.
Is it possible to have this kind of staining? Or do there have any suggestion to improve better nuclear staining of the antibody for IHC frozen section?