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SantaCruz Antibody For Two Hybrid Assay (Again)

Marian Waterman mlwaterm at uci.edu
Mon Feb 13 16:48:44 EST 1995


Shun' Ichi

   We use Santa Cruz's Gal4 antibody to detect Gal4 protein in our
two-hybrid strains.  We use a 1/1000 dilution of the primary antibody, and
a 1/5000 dilution of a mouse secondary antibody. Gal4 production from a
pAS vector is extremely high and readily detectable in one to two seconds
using ECL detection.  Our yeast extracts are made in RIPA buffer and lysed
with glass beads.  We are loading the equivalent of about 2 mls of yeast
(OD = 0.5) per one lane on a protein mini-gel.

If this is not clear, please feel free to contact me.  I do not scan this
newsgroup regularly.

Sincerely,
Marian Waterman
Microbiology and Molecular Genetics
UC Irvine, CA
mlwaterm at uci.edu



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