>Hi:
>.I have been having problems getting antibody from ascites fluid
>In the procedure I used, Balb/C mice (female) were primed with
>pristane (0.25 ml/mouse). After 15 d 3million hybridoma cells were
>injected per mouse (in 0.5 ml). In about 2 weeks, the mice began to
>show swelling, with some producing solid tumors and others producing
>ascites fluid. However, RIA of the ascites fluid (or of a 0-50%
>ammonium sulfate cut of the fluid) did not reveal the presence of the
>antibody.
>Any insight/suggestions regarding this problem would be appreciated.
>Thanks,
>Tilak Ponappa (tkp2 at po.cwru.edu
Hi
It happens.
1. Your hybridoma might be unstable
2. This particular mAB specificity might be toxic so that only
unproducing/mutated clones survive only
3. Highest titers are obtained from mice killed just before their "natural"
death though it might sound cruel to sit and watch and wait...
4. Solid tumors (in part of your animals) indicate that something is
wrong with the host. Are you getting them from a sure source?
If your hybridoma is well cloned and stable in culture BEFORE
going into mice I would try to culture some cells from ascite
and see if they start to produce AB again
Best wishes
Andrei
