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Having incubated the immunoprecipitated samples overnight at 4 C with rotation, the anti-M2 FLAG antibody-conjugated agarose beads were washed three times (each for 5 min) with ice-cold washing buffer (50 mM Tris, pH 7.4, 137 mM NaCl, 10% glycerol, 1% NP-40)

while effective, it may offer slightly reduced benefits compared to IV administration

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It is often recommended to include a prior washing step, such as rinsing culture plates with cold PBS immediately before quenching, to minimize carryover effects

Eur Rev Med Pharmacol Sci (2021) 25(4):182836