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After blocking, tumor cells were stained with APC-Cy7 conjugated anti-mouse CD45 (BD biosciences, Cat # 557659), BV711 conjugated anti-mouse CD3 (BioLegend, Cat # 100241), PE conjugated anti-mouse NKp46 (BD biosciences, Cat # 560757), AF488 conjugated anti-mouse CD11c (BioLegend, Cat # 117313), and BV421 conjugated anti-mouse CD11b (BioLegend, Cat # 101235)
Since the aim was to generate a pFastBac1 allowing the expression of recombinant protein with streptavidin- and 10xHis-tags, the BamHI restriction site of the pET-52b(+) MCS sequence was not used
However, the reliability of this technique is undermined by the formation of GST-fused protein aggregates after incubation with cell lysates
In other words, NAD+ is a key component of the cellular energy generating system
