Detecting b-gal in cultured cells ?

Dave Bates dobates at
Fri Jun 10 14:33:04 EST 1994

In article <simmenlab.33.2DF78727 at>
simmenlab at writes:

> Hello:
>               I am transfecting cells in culture with a b-gal expression 
> vector.  I was wondering if it was possible to detect b-gal expression without 
> preparing cell extracts.  I just want to know if the transfection worked or 
> did not work.  If anyone has any experience with this please let me know.  
> Thank you very much!
>                                                           Karen Reed

For cultured endothelial cells I wash with PBS,  fix with
2%Formaldehyde, 0.1%Glut, Wash again and stain O/N at 37C with X-Gal
assay buffer (60uls 50mg/ml X-Gal, 75ul each of  K Ferro and
Ferricyanide (0.2M), 12uls MgCl2 (1M), made up to 3mls with PBS).
Worksd a treat. Can also use anti B gal antibodies (from lots of places
eg Sigma), or for in vivo work without fixation, Image green or Imagene
Red from Molecular Probes.

Good Luck
Dave Bates,                                        dobates at
Department of Human Physiology, University of California at Davis
No I don't have a sense of humour, now buy me a beer

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