probing for pAMPK:
blocking - 5% bsa
primary - 2% bsa o/n
secondary - 2% bsa
washings with 0.1% tbst
my queries are:
should i be blocking with nfdm?
and secondary also with nfdm?
and for total ampk
should continue with bsa?
or with milk for blocking /primary/secondary?
Elizabeth C. Hinchy
4% v/v Odyssey Blocking Buffer in TBST works well for all antibody incubations, particularly if you are detecting secondaries by a fluorescent method. But the other blocking agents mentioned (diluted in TBST) should also work. There should be no need to switch between blocking agents for primary and secondary antibody incubations.
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