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Tes overnight at 4 . For chain, IgG1, IgG2c isotype determination, sera Tes overnight at 4 . For chain, IgG1, IgG2c isotype determination, sera were diluted 1/500, 1/5000, and 1/1000, respectively, and added to the wells for overnight incubation at 4 . Biotin-conjugated https://britishrestaurantawards.org/members/debt31yacht/activity/286127/ antibodies specific for chain, IgG1, IgG2c were added and incubated for 1 hr at 37 , followed by strepavidin-alkaline conjugate according to the manufacturer's recommendations (BD Pharmingen). Experimental chronic GVH protocol Chronic GVH was induced as previously described [24]. Briefly, donor bm12 splenocytes (1?08) were injected i.p. into TRAF1KO or WT recipient mice at the age of 12?6 weeks. Mice were bled serially for the following 9 weeks, and anti-chromatin antibodies were quantitated as.