Hello everyone,

recently i am performing immunofluorescence analysis experiments and i came across the anti-HAS2 goat polyclonal antibody (S-15, sc-34067) from santa cruz biotechnology, but i am not able to detect proper staining. I only observe weak nuclear staining and diffusion , i have also tried in a different cell line and the problem remained. The protocol i usually use is this : Fixation 4% PFA, Permeabilization : 0.05% Triton X-100, Blocking : 5% BSA, 1st Ab: 1:200 in 1% BSA/PBS-Tween (also tried 1:100), 2nd Ab: tried many dilutions in 1% BSA/PBS-Tween (for 1 hour), Washes : 0.01% PBS-Tween. Have anyone used this antibody? Can anyone suggest a different protocol? any ideas? 

Thanks in advance