iNOS and nNOS protein quantification from rat tissue homogenate - (Jun/05/2006 )

Hi There,

I am looking for some protocools for non-radioactive quantification of iNOS and nNOS protein from rat tissue homogenate.

Anybody have tried that. I will be greatfull if any body help me. I am prefering either ELISA or WB.

Thanks in adavance.

Hi,

I have been doing Westerns looking for all the NOS isoforms from rat prostate cancer cells.

iNOS and nNOS have been quite successful using Santa Cruz antibodies.
I use a 7.5 % gel, transfer over night, block with 5 % milk and then add primary antibody (both at 1:100 diluted in 5% milk) then 3 x 10 mins washes in PBS/tween and secondary of swine anti-rabbit from Dako at 1:3000, then 3 more washes.

Hope that helps a little.