i attached the picture and have a some problem.
i stained EEA1(early endosome marker). however there are many spots in the nuclei. it shouldn't be observed i think.
is it normal? or should i change some protocol?
this is my short protocol
used -20 degree methanol for fixation and permeablization 10minute.
blocked with 3%BSA/PBS 30minute RT
stained EEA1 diluted in 3%BSA/PBS 1hour RT with low speed shaking.
wash 3times PBS and confocal microscopy
please reply for my topic for my work. HHHHELP Me~~~
Edited by flightyoon, 18 June 2014 - 01:22 AM.