Memory B cells were differentiated in vitro into plasmablasts and plasma cells, following the protocol described in Jourdan et al. (24 (link)). Briefly, CD27+ Memory B cells were first purified as bulk B cells by negative selection from PBMCs by magnetic isolation followed by positive selection for CD27+ cells (Stemcell Technologies, UK) following manufacture's protocol. Then seeded at1.5 × 105 cells/ml, and cultured for 4 days in the presence of CpG oligodeoxynucleotide 2006 (10 μg/ml; Invitrogen), soluble human his-rCD40L (50ng/ml; R&D Systems, UK) and anti-poly-his (5μ g/ml; R&D Systems, UK), IL-2 (20 U/ml; R&D Systems, UK), IL-10 (50 ng/ml; Miltenyi Biotec, UK) and IL-15 (10 ng/ml; Miltenyi Biotec, UK). At day 4 of culture, the cells were washed and cultured at 2.5 × 105 cells/ml with IL-2 (20 U/ml, Milteny Biotec, UK), IL-10 (50 ng/ml), IL-15 (10 ng/m, PreproTech, UK) and IL-6 (50ng/ml; Miltenyi Biotec, UK). At day 7 of culture, cells were washed and seeded with IL-6 (50 ng/ml, PreproTech, UK), IL-15 (10 ng/ml, PreproTech, UK) and IFN-α (500 U/ml; R&D Systems) for 3 days.
Anti poly his
Manufactured by R&D Systems
Sourced in United Kingdom
Anti-poly-his is a lab equipment product designed to detect and quantify poly-histidine tagged proteins. It functions as an antibody that specifically binds to the poly-histidine tag, allowing for the identification and isolation of recombinant proteins labeled with this tag.
Automatically generated - may contain errors
Lab products found in correlation
Cd27 cells, by STEMCELL (2 mentions)
Soluble human his rcd40l, by R&D Systems (2 mentions)
Cpg oligodeoxynucleotide 2006, by Thermo Fisher Scientific (2 mentions)
Il 10, by Miltenyi Biotec (2 mentions)
Il 10, by Thermo Fisher Scientific (2 mentions)
Il 15, by Thermo Fisher Scientific (2 mentions)
Interleukin 6 (il 6), by Miltenyi Biotec (2 mentions)
Interleukin 6 (il 6), by Thermo Fisher Scientific (2 mentions)
Ifn α, by R&D Systems (2 mentions)
Interleukin 2 (il 2), by Miltenyi Biotec (2 mentions)
Il 15, by Miltenyi Biotec (2 mentions)
Interleukin 2 (il 2), by R&D Systems (2 mentions)
2 protocols using anti poly his
1
Differentiation of Memory B Cells into Antibody-Secreting Cells
2
In Vitro Differentiation of Memory B Cells
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Memory B cells were differentiated in vitro into plasmablasts and plasma cells, following the protocol described in Jourdan et al. (24 (link)). Briefly, CD27+ Memory B cells were first purified as bulk B cells by negative selection from PBMCs by magnetic isolation followed by positive selection for CD27+ cells (Stemcell Technologies) following the manufacturer’s protocol. The cells were then seeded at 1.5 × 105 cells/ml and cultured for 4 d in the presence of CpG oligodeoxynucleotide 2006 (10 μg/ml; Invitrogen, Paisley, U.K.), soluble human his-rCD40L (50 ng/ml; R&D Systems, Abingdon, U.K.) and anti–poly-his (5 μg/ml; R&D Systems), IL-2 (20 U/ml; R&D Systems), IL-10 (50 ng/ml; Miltenyi Biotec, Surrey, U.K.), and IL-15 (10 ng/ml; Miltenyi Biotec). At day 4 of culture, the cells were washed and cultured at 2.5 × 105 cells/ml with IL-2 (20 U/ml; Miltenyi Biotec), IL-10 (50 ng/ml), IL-15 (10 ng/ml; PeproTech, London, U.K.), and IL-6 (50 ng/ml; Miltenyi Biotec). At day 7 of culture, cells were washed and seeded with IL-6 (50 ng/ml; PeproTech), IL-15 (10 ng/ml; PeproTech), and IFN-α (500 U/ml; R&D Systems) for 3 d.
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