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Based on the determined protein concentration, 100 g of protein solution was taken, and the volume was adjusted to 200 L with 8 M urea

Unbound antibodies were washed off by rocking and rinsing the slices thrice with 0.1 M PBS, followed by 2 h of incubation at room temperature with corresponding fluorescence-tagged secondary antibodies (diluted in blocking solution)

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doi: 10.1002/jimd.12281