OKT 8 antibody - (May/22/2011 )

I am trying to purify OKT8 antibody from mouse ascites. I tried in balb/c mouse but ascites didn't develop. can any body trouble shoot me. how it can be done? also how OKT8 hybridoma has been generated? what strategy has been used to confirm them?

thanks

Hi!
You must be sure your hybridoma is syngeneic with the Balb/c mouse strain. In other words, lymphocytes and myeloma cells used for the fusion have to be Balb/c cells as well. Otherwise, the hybridoma cells might be rejected by the mouse immune system.
If your hybridoma is actually syngeneic, make sure you inject pristane the mice.
I hope this helps.

pristane i have used. Mouse is not syngenic, but its not even rejected . there is no immune response symptoms

Chelo on Sun May 22 17:52:29 2011 said:

shilpeejnc on Mon May 23 12:24:05 2011 said:

Hi again!

I am not sure to follow you arguments. If cells are rejected, you won´t see any "symptoms" since we are talking about individual cells, not an organ. In other words, your hybridoma cells must be killed and you won´t be able to notice that. Better change mouse strain. Alternatively, try higher number of injected cells. We try with 1 million cells for isogenic injection. You might want to try 10-100 times that number for non-related mice.