detecting His-tagged gene products

Theo Hanekamp hanekamp at uwyo.edu
Mon Feb 16 17:32:26 EST 1998


We have used PCR to HIS-tag the 3' end of three different genes in
Saccharomyces cerevisiae.  These His-tagged fusions complement the mutant
phenotype when expressed in yeast strains carrying the appropriate null
allele. Furthermore, DNA sequence data show that the HIS-tag has been
added correctly. However, we have not been able to detect these
His-tagged fusions on Western blots using the INDIA-HIS probe (from
Pierce), although we religiously followed the protocol that was included
in the INDIA HIS probe Western kit.  Has anyone used the INDIA HIS probe
from Pierce and got it to work in yeast? Has anyone successfully used
other anti-HIS antibodies in yeast?  Suggestions/ comments are greatly
appreciated. Please, send your response to:  hanekamp at uwyo.edu 

Thank you very much in advance,
Theo



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