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Neutrophils were isolated from peripheral blood using HiSep 1077 and GranuloSep GSM 1119. Initially, a double gradient (Percoll density gradient) is formed by layering an equal volume of HiSep 1077 over GranuloSep GSM 1119. Blood is carefully layered on to the upper HiSep 1077 medium and centrifuged (3,000 rpm for 30 min.) at room temperature. Neutrophils present at the 1077/1119 interphase layer were isolated, washed twice in RPMI 1,640 medium, centrifuged (1,600 rpm for 10 min.), and suspended in RPMI 1640. We checked cell viability using trypan blue (>95%).
Copyright and License information: ©2021 Journal of Family Medicine and Primary Care
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