Why are my B-hybrid cells dying?

[ZIMMORN]

Hi Everyone,

I am making monoclonal antibodies now. I screened all the hybrid clones by ELISA and picked up those positive ones, which are then sub cloned by limited dilution. One of the clones initially grew pretty well and is dying now. Under the microscope, I found the clones probably have 50-100 cells, but all of them has ugly shapes, indicating they are dead.

Why are these cell dying? And any suggestion for rescuing them?

Thanks in advance.

Zimmorn

[MicroKiller]

hi there.. not too sure but i did encounter the same prob last time. I jst went back to the original stock of the plate (hopefully u made a frozen stock) and did the dilution again but this time with lower dilutions (its more work) but it ensures the cells survive the passaging.

A crazy idea.. but would it be possible to percipitate the cells and inject them back into a mice to generate ascites fluid.. ??