Hi, I have some flash-frozen mouse spleens (in the -80C) that I'd like to analyze for CD3, preferably using an Elisa kit to reduce time. I've been searching various sites, but not a lot of options are out there for tissue (most are targeted at plasma). Obviously, using flow cytometry is out because the spleens are frozen.

Also, we didn't extract blood out before freezing - Do you have a protocol you can share that we could use to get rid of the excess RBCs after thawing? Would thaw/cut/rinse with chilled PBS still work, or would using a standard tissue lysis buffer (Tris-based) with protease inhibitor do the trick since we will be centrifuging and taking off supernatant in any case?

Any recommendations for protocols, products/antibodies/Elisas or companies would be greatly appreciated.