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After primary antibody incubation, the cells were washed 4 times with PBS-T and incubated with fluorophore labeled secondary antibodies diluted in PBS for 1 hour at room temperature in the dark

Less acid means less available B12

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However, no manufacturer or regulatory body recommends using it past the expiration date, and most healthcare providers advise against it because the actual potency in your specific vial is unknowable without laboratory testing