Hi,
I've just started staining with mouse mAb's against human VCAM-1 and ICAM-1 (Santa Cruz) but have not had any luck... I'm using paraffin embedded human biopsies and the protocol(which is otherwise standard) includes citrate antigen retrieval, blocking in 2% secondary Ab species serum for 1h @RT, incubation with the primary Ab overnight @4 deg. C at dilutions ranging from 1:50 to 1:100. I have also tried DAB times up to 10 min. The above protocol works very well normally but for these antibodies I just can't seem to get any staining. Maybe I'm just thick but if anyone has any suggestions as to what may be the problem, please let me know!
Cheers,
