LacZ staining on tissue extracts

gtk10583 gtk10583 at glaxowellcome.com
Fri Sep 21 13:06:36 EST 2001


I am interested in staining for LacZ in tissues from a transgenic mouse
overexpressing a b-gal transgene.

I have tried mincing the tissue in a digest buffer containing triton x-100
and clearing the extract via centrifugation.  However, the assay I am using
for b-gal expression is a colorimetric assay using ONPG as substrate.  The
problem is most tissue extracts are heavily pigmented and mask the color
generated by enzymatic activity.  Is there a better protocol for extracting
protein from whole tissues for lacZ staining?




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