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A Loss of Affinity Mutants (LAM) screening yields interaction-deficient point mutants of your bait protein for a given partner. LAM takes advantage of our ULTImate Y2H™ technique.
Use LAM screening to map interaction hotspots and obtain dominant negatives to speed up your functional studies!
CONTACT US and submit the sequences of your proteins.
Mutants are generated by PCR and the experimental conditions are adapted to each target gene to get on average a single mutation per clone.
The mutant library is then screened against the interacting partner by yeast two-hybrid using the LacZ reporter gene. White or light blue yeast clones in which no or a weak interaction occurs are selected.
192 clones are fully sequenced to identify their mutation. Their phenotypes are confirmed in a secondary, semi-quantitative LacZ assay.
You select up to 10 point mutants which are further characterized in a quantitative colorimetric interaction assay. You then receive DNA plasmids for these 10 mutants for your own susbsequent functional studies.