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“…Antibody staining was performed at dilution 1 : 500 as previously described by Rougon et al [36,38]. Nonspecific binding was reduced by preincubating the antisera with the KLH carrier (1 mM).…”
Section: Resultsmentioning
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“…Antibody staining was performed at dilution 1 : 500 as previously described by Rougon et al [36,38]. Nonspecific binding was reduced by preincubating the antisera with the KLH carrier (1 mM).…”
Section: Resultsmentioning
“…Peptides were coupled to keyhole limpet hemocyanin (KLH) with either glutaraldehyde or N-ethyl, N' dimethylaminopropyl carbodiimide (ECCD) as described previously by Rougon et al [36,38]. In order to orient the coupling of peptide 1 towards the carboxyl-terminus, amino, hydroxy and sulfhydryl groups were protected during coupling reactions by a reversible citraconic acid modification [8].…”
Section: Production Of Antibodiesmentioning
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“…All peptides were C-terminally amidated with free amino termini. Each peptide was coupled to bovine serum albumin (BSA) [21] to promote binding of the peptide to the ELISA plate.…”
Section: Methodsmentioning
“…W6\32 hybridoma was also injected into mice for ascites production (Harlow & Lane, 1988). Rabbit antiserum specific for a peptide sequence on the cytoplasmic tail of human MHC class I heavy chain (Rougon et al, 1984), and denatured HLA-B27 (Neefjes et al, 1986) were generous gifts from H. L. Ploegh.…”
Section: Methodsmentioning