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Instead of relying on endpoint determination, it identifies early growth inflection points and establishes a combined interpretation system of early bacterial growth kinetics + PPT threshold + slope verification, effectively reducing the risk of misjudgment

For intracellular cytokine, staining cells were fixed with BD Cytofix Cytoperm (BD Biosciences) for 20 min at 4 C, washed twice with 1x saponin buffer (0.5% (w/v) saponin [Carl Roth], 2 mM EDTA, 2% FCS, in PBS) and stained for IFN (see Table 1) overnight at 4 C in 1 saponin buffer

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All the data comes from one group, so people can be skeptical

Indeed, because more than 100 proteins have been reported as TG2 substrates and because TG2 has broad substrate specificity, the GST tag could be crosslinked with a number of cellular proteins 21