Signal in the citoplasm instead of in the nucleus - (Nov/30/2008 )
Hello!!
I am pretty new in inmunohistochemistry so if someone here can give me a hand would be great. This is my problem: I am trying two ab against Pit-1 following the recomendations of the house(aviva systems) an using a positive control than I have checked it is really positive. But these ab signal the citoplasm while they should singal the nucleus. How can I solve this? My protocol includes microwave ph6, peroxidase, streptavidine- biotine and DAB. I also tried different dilutions and time of incubation but it keeps on signaling the citoplasm
Thanks in advance!!
-Marinla-
QUOTE (Marinla @ Nov 30 2008, 04:59 AM)
Hello!!
I am pretty new in inmunohistochemistry so if someone here can give me a hand would be great. This is my problem: I am trying two ab against Pit-1 following the recomendations of the house(aviva systems) an using a positive control than I have checked it is really positive. But these ab signal the citoplasm while they should singal the nucleus. How can I solve this? My protocol includes microwave ph6, peroxidase, streptavidine- biotine and DAB. I also tried different dilutions and time of incubation but it keeps on signaling the citoplasm
Thanks in advance!!
I am pretty new in inmunohistochemistry so if someone here can give me a hand would be great. This is my problem: I am trying two ab against Pit-1 following the recomendations of the house(aviva systems) an using a positive control than I have checked it is really positive. But these ab signal the citoplasm while they should singal the nucleus. How can I solve this? My protocol includes microwave ph6, peroxidase, streptavidine- biotine and DAB. I also tried different dilutions and time of incubation but it keeps on signaling the citoplasm
Thanks in advance!!
attach photos, we might be able to help. but don't forget cells are in 3D, not 2D, so still you might see some light coming from the nucleus, but it might not necessarily be from nucleus, it might be from the cytoplasm above the nucleus.
-Curtis-