>What are the considerations when tagging proteins for
immunofluorescence or pur
>ification of complexes?
For immunoflourescence, check that the tagged protein WHEN EXPRESSED AT
WILD TYPE LEVELS can functionally complement a full disruption of the
gene in question. If you must overexpress the tagged protein for
complementation, you will never be assured that the gene product is
properly localized. I would suggest that the safest way to check this
is to replace the GENOMIC wild type gene with the tagged gene to insure
proper levels of expression.
The most common places to tag proteins is right on the N- or C-terminus
of the protein. Hope this helps, Morrie
******************************************************************************
Morris F. Manolson Tel: 416-813-6662 (office)
Division of Cell Biology 416-813-5594 (lab)
Hospital for Sick Children 416-813-5028 (FAX)
88 Elm St., McMaster building email: Morrie at resunix.ri.sickkids.on.ca
Toronto, Ontario, Canada
M5G 1X8
