Dear all, I have been experiencing a problem with PCNA flow staining on monkey peripheral blood monocytes. To be able to label nuclear antigens such PCNA, I have used perm & fix steps (Triton- and methanol-based) prior to antibody incubation. The thing is, after methanol fixation my cells are gone. I am asking for a piece of wisdom. Outline of the protocol I have used is following: 1. Surface antigen staining for whole blood 2. Red blood cell lysis 3. 2% paraformaldehyde staining 4. 5% Triton X-100 for 15 min 5. 100% methanol at -20 degree for 15 min 6. PCNA staining. I would very much appreciate if any of you helped me troubleshoot this problem. Thank you. Cheers, Woong-Ki Kim Woong-Ki Kim, Ph.D. Beth Israel Deaconess Medical Center 330 Brookline Ave RE 113 Boston, MA 02215 E-mail: wkim1@caregroup.harvard.edu
This archive was generated by hypermail 2b29 : Sun Jan 05 2003 - 19:26:25 EST