Methods and Techniques Discussion => Immunocytochemistry (ICC) => Topic started by: MariaKokki on July 27, 2007, 02:34:35 PM

Title: BrdU staining and trypsin pre-treatment on cells
Post by: MariaKokki on July 27, 2007, 02:34:35 PM
Hi! I want to label a cell line with BrdU and then stain the cells grown on coverslips with anti-BrdU. I will use DAB chromate for staining. According to my antibody's protocol, I need to add trypsin (0.1% in PBS for 20min at 37oC !) before the incubation with BrdU.
Does anybody have any idea of why is that? I am afraid that my cells will detach from the coverslip. Is trypsin pre-treatment really necessary in my case?
Title: Re: BrdU staining and trypsin pre-treatment on cells
Post by: excalibur on July 27, 2007, 06:20:12 PM
As always, you will just need to try it and see. But cells can be fixed for shorter times than tissues, so trypsin may or maynot be needed. Have you spoken with tech support of the compnay that made your antibody?