Author: Bancroft, Tara; DeBuysscher, Blair L.; Weidle, Connor; Schwartz, Allison; Wall, Abigail; Gray, Matthew D.; Feng, Junli; Steach, Holly R.; Fitzpatrick, Kristin S.; Gewe, Mesfin M.; Skog, Patrick D.; Doyle-Cooper, Colleen; Ota, Takayuki; Strong, Roland K.; Nemazee, David; Pancera, Marie; Stamatatos, Leonidas; McGuire, Andrew T.; Taylor, Justin J.
Title: Detection and activation of HIV broadly neutralizing antibody precursor B cells using anti-idiotypes Document date: 2019_10_7
ID: 63yvpuqx_46
Snippet: After centrifugation, supernatant was discarded and cell pellets were resuspended in FACS buffer. The entire enriched fraction and 1/40th of the flow-through or unenriched fractions and were incubated with antibodies against surface markers in a total volume of 50 µl for 25 min on ice and washed with FACS buffer. Human PBMCs were labeled with 6.24 µg/ml anti-IgM FITC (G20-127; BD Biosciences), 4 µg/ml anti-IgD PerCP-Cy5.5 (IA6-2; BD Bioscience.....
Document: After centrifugation, supernatant was discarded and cell pellets were resuspended in FACS buffer. The entire enriched fraction and 1/40th of the flow-through or unenriched fractions and were incubated with antibodies against surface markers in a total volume of 50 µl for 25 min on ice and washed with FACS buffer. Human PBMCs were labeled with 6.24 µg/ml anti-IgM FITC (G20-127; BD Biosciences), 4 µg/ml anti-IgD PerCP-Cy5.5 (IA6-2; BD Biosciences), 2 µg/ml anti-CD20 eFluor 450 (2H7; eBioscience), 1 µl anti-CD27 BV650 or BV480 (L128; BD Biosciences), 1 µl anti-CD3 BV711 (UCT1; BD Biosciences), 1 µl anti-CD14 BV711 (MϕP9; BD Biosciences), 2.8 µg/ml anti-CD16 BV711 (3G8; BioLegend), and 1 µl anti-CD19 PE-Cy7 (HIB19; BD Biosciences).
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