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The 72-hour threshold is not an official manufacturer guideline it's a practical working limit based on how quickly bacterial growth can become concerning at typical room temperatures (2025C) even in the presence of benzyl alcohol

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The membranes were blocked for 1 h at room temperature in 5% non-fat dry milk (4259.01, SERVA, Heidelberg, Germany) prepared in Tris-buffered saline with Tween-20 (P7949, Sigma-Aldrich, Pozna, Poland) (TBST)