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FeSO 4 solutions of varying concentrations (0, 0.01, 0.025, 0.05, 0.1 ppm) were prepared in 1% Tween20 and applied on L

It should be kept away from strong chelators (such as high levels of EDTA and related sequestrants), reducing sugars, thiol-bearing actives, and high concentrations of direct acids (for example ascorbic acid or alpha-hydroxy acids at low pH), all of which can compete for the copper center or disrupt the peptide-metal bond and shift the characteristic blue color toward brown or colorless degradation products

The resultant single cells were stained with antibodies purchased from Biolegend (USA): PE/Cy7 anti-mouse CD45 (1:400, #103113), FITC anti-mouse CD3 (1:400, #100203), APC anti-mouse CD4 (1:400, #100411), PE anti-mouse CD8 (1:400, #100707), APC anti-mouse IFN-(1:100, #505809)