LacZ staining on tissue extracts

Sergio Alonso sergioal at bbm1.ucm.es
Mon Sep 24 06:34:54 EST 2001


maybe a protocol based on X-Gal inestead of ONPG???

gtk10583 ha escrito:

> 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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